Aromatic amidine derivatives and salts thereof

ABSTRACT

An anticoagulant agent which comprises, as an active ingredient, an aromatic amidine derivative represented by the following general formula (1) or a salt thereof: ##STR1## wherein the group represented by ##STR2## is a group selected from indolyl, benzofuranyl, benzothienyl, benzimidazolyl, benzoxazolyl, benzothiazolyl, naphthyl, tetrahydronaphthyl and indanyl; X is a single bond, an oxygen atom, a sulfur atom or a carbonyl group; and Y is a saturated or unsaturated 5- or 6-membered heterocyclic moiety or cyclic hydrocarbon moiety optionally having a substituent group, an amino group optionally having a substituent group or an aminoalkyl group optionally having a substituent group. 
     The inventive compound has a high anticoagulant capacity based on its excellent FXa inhibition activity.

This is a continuation of parent application Ser. No. 08/282,571, filedJul. 29, 1994; now abandoned which is a continuation of application Ser.No. 07/969,396, filed Oct. 30, 1992, now abandoned.

FIELD OF THE INVENTION

This invention relates to aromatic amidine derivatives and saltsthereof, which are capable of showing a strong anticoagulant effectthrough reversible inhibition of activated blood coagulation factor X(hereinafter, referred to as "FXa") and can be administered orally. Theinvention also relates to an anticoagulant, or a thrombosis- orembolism-preventing or treating agent that contains the aromatic amidinederivative or a salt thereof as an active ingredient.

BACKGROUND OF THE INVENTION

Attempts have been made in the prior art to develop an antithrombinagent as an antithrombotic agent. However, it has been known that suchan antithrombin agent is apt to cause bleeding tendency and difficultyto manage haemostasis because it inhibits blood coagulation and alsothrombin induced platelet aggregation. With the aim of overcoming suchproblems, the development of anticoagulant agents has been attemptedbased on a inhibitory mechanism other than the thrombin inhibition. As aresult of such efforts, 1,2-bis(5-amidino-2-benzofuranyl)ethane(hereinafter, referred to as "DABE") represented by the followingformula (2) has been found as an anticoagulant agent based on FXainhibition (Thrombosis Research, vol.19, pp. 339-349, 1980): ##STR3##

However, DABE has the disadvantages that it has both FXa and thrombininhibitory activities which cannot be separated sufficiently from eachother, it has a very low water solubility, and it does not show itsanticoagulant effect when administered orally. Consequently, greatattention has been directed, from a clinical point of view, toward thedevelopment of a drug which is highly specific and potent inhibitor forFXa, has a high water solubility, and is effective in oraladministration.

SUMMARY OF THE INVENTION

In view of the above, the present inventors have conducted intensivestudies on the synthesis of various types of aromatic amidinederivatives and evaluated their pharmacological properties. As a resultof such efforts, they have found that an aromatic amidine derivativerepresented by the following general formula (1) or a salt thereofpossesses excellent water solubility, shows a strong anticoaguianteffect through its highly specific and reversible FXa-inhibitingactivity even in the case of oral administration, and is useful as adrug for the prevention and treatment of various thrombosis- andembolism-based diseases. The present invention has been accomplished onthe basis of these findings.

That is, according to the present invention, there is provided anaromatic amidine derivative represented by the following general formula(1) or a pharmaceutically acceptable salt thereof: ##STR4## wherein R¹is a hydrogen atom or a lower alkoxy group; R² is a hydrogen atom, alower alky group, a lower alkoxy group, a carboxyl group, analkoxycarbonyl group, a carboxyalkyl group or an alkoxycarbonylalkylgroup; R³ is a hydrogen atom, a carboxyl group, an alkoxycarbonyl group,a carboxyalkyl group, an alkoxycarbonylalkyl group, a carboxyalkoxygroup or an alkoxycarbonylalkoxy group; R⁴ is a hydrogen atom, ahydroxyl group, a lower alkyl group or a lower alkoxy group; n is aninteger of 0 to 4; A is an alkylene group having a carbon number of 1 to4, which may have 1 or 2 substituents selected from the group consistingof hydroxyalkyl, carboxyl, alkoxycarbonyl, carboxyalkyl andalkoxycarbonylalkyl; X is a single bond, an oxygen atom, a sulfur atomor a carbonyl group; Y is a saturated or unsaturated 5- or 6-memberedheterocyclic moiety or cyclic hydrocarbon moiety optionally having asubstituent, an amino group optionally having a substituent, or anaminoalkyl group optionally having a substituent; and the grouprepresented by ##STR5## is a member selected from the group consistingof indolyl, benzofuranyl, benzothienyl, benzimidazolyl, benzoxazolyl,benzothiazolyl, naphthyl, tetrahydronaphthyl and indanyl, preferably anindolyl, benzofuranyl, benzothienyl, benzimidazolyl, benzothiazolyl,naphthyl or tetrahydronaphthyl group, more preferably an indolyl,benzothienyl or naphthyl group.

The present invention also provides an anti-coagulation agent, or athrombosis- or embolism-preventing or treating agent which contains acompound of general formula (1) or a salt thereof as an activeingredient.

DETAILED DESCRIPTION OF THE INVENTION

In the compound of the present invention represented by general formula(1), any straight chain, branched chain or cyclic alkyl group having 1to 6 carbon atoms may be used as the lower alkyl group. Illustrativeexamples include methyl, ethyl, propyl, isopropyl, butyl, sec- ortert-butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl and the like. The lower alkoxy group may have 1 to 6 carbonatoms. Illustrative examples include methoxy, ethoxy, propoxy,isopropoxy, butoxy, sec- or tert-butoxy and the like. Thealkoxycarbonyl, carboxyalkyl, alkoxycarbonylalkyl, carboxyalkoxy,alkoxycarbonylalkoxy and hydroxyalkyl groups preferably have 1 to 6carbon atoms, more preferably 1 to 4 carbon atoms, respectively.Illustrative examples of the alkoxycarbonyl group includemethoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl and thelike. Illustrative examples of the carboxyalkyl group includecarboxymethyl, carboxyethyl, carboxypropyl and the like. Illustrativeexamples of the alkoxycarbonylalkyl group include methoxycarbonylmethyl,ethoxycarbonylmethyl, propoxycarbonylmethyl, methoxycarbonylethyl,ethoxycarbonylethyl, methoxycarbonylpropyl, ethoxycarbonylpropyl and thelike. Illustrative examples of the carboxyalkoxy group includecarboxymethoxy, carboxyethoxy, carboxypropoxy and the like. Illustrativeexamples of the alkoxycarbonylalkoxy group includemethoxycarbonylmethoxy, ethoxycarbonylmethoxy, propoxycarbonylmethoxy,methoxycarbonylethoxy, ethoxycarbonylethoxy and the like. Illustrativeexamples of the hydroxyalkyl group include hydroxymethyl, hydroxyethyl,hydroxypropyl, hydroxybutyl and the like. Illustrative examples of thealkylene group having 1 to 4 carbon atoms and represented by A includemethylene, ethylene, trimethylene, tetramethylene and the like.

The saturated or unsaturated 5- or 6-membered heterocyclic moiety maycontain preferably one or two hetero-atom(s) selected from nitrogen andoxygen atoms. Illustrative examples of such a preferred type ofheterocyclic rings include pyrrolidine, piperidine, imidazoline,piperazine, tetrahydrofuran, hexahydropyrimidine, pyrrole, imidazole,pyrazine, pyrrolidinone, piperidinone, morpholine and the like. Morepreferable are pyrrolidine and piperidine which contain one nitrogenatom as the hetero-atom. Illustrative examples of the saturated orunsaturated cyclic hydrocarbon moiety include cyclopentyl, cyclohexyland the like. Illustrative examples of the aminoalkyl group includeaminomethyl, aminoethyl, aminopropyl and the like. Illustrative examplesof the substituents applicable to these heterocyclic moieties and cyclichydrocarbon moieties include preferably lower alkyl, lower alkanoyl,carbamoyl, mono- or dialkylcarbamoyl, formimidoyl, alkanoimidoyl,benzimidoyl, carboxyl, alkoxycarbonyl, carboxyalkyl, alkylcarbonylalkyl,aminoalkyl, alkanoylamino, alkanoylaminoalkyl, imino,alkoxycarbonylimino and the like, more preferably formimidoyl andalkanoimidoyl groups. Illustrative examples of the substituentsapplicable to these amino groups and amino moieties of aminoalkyl groupsinclude preferably lower alkyl, pyradinyl, pyrrolidinyl, carbamoyl,mono- or dialkylcarbamoyl, lower alkanoyl, formimidoyl, alkanoimidoyl,benzimidoyl, alkoxycarbonyl and the like, more preferably pyrazinyl,pyrrolidinyl, formimidoyl, alkanoimidoyl groups. In this instance, eachof the alkyl, alkoxy, alkanoyl and the like listed above may preferablyhave a carbon number of from 1 to 6.

The compound of formula (1) of the present invention may have an opticalisomerism or a stereoisomerism due to the presence of an asymmetriccarbon atom. An optical isomer, a stereoisomer and a mixture thereof areincluded in the scope of the present invention.

Salts of the compound of formula (1) of the present invention are notparticularly limited, provided that they are pharmaceuticallyacceptable. Illustrative examples of such salts include: inorganic acidsalts such as hydrochloride, hydrobromide, hydroiodide, phosphate,nitrate, sulfate and the like; organic sulfonic acid salts such asmethane sulfonate, 2-hydroxyethane sulfonate, p-toluene sulfonate andthe like; and organic carboxylic acid salts such as acetate, propanoate,oxalate, malonate, succinate, glutarate, adipate, tartarate, maleate,malate, mandelate and the like.

Most preferred examples of the compound of formula (1) of the presentinvention are as follows:

2-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid;

(+)-2-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid;

(2S)-2-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid;

(2R)-2-[4-[((3R)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid;

2-[4-[(1-acetimidoyl-4-piperidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid;

(+)-2-[4-[(1-acetimidoyl-4-piperidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid;

2-[4-[(1-acetimidoyl-4-piperidinyl)oxy]phenyl]-3-(5-amidinobenzo[b]thien-2-yl)propionicacid;

2-[4-[((2S)-1-acetimidoyl-2-pyrrolidinyl)methoxy]phenyl]-3-(5-amidinobenzo[b]thien-2-yl)propionicacid;

(+)-2-[4-[((2S)-1-acetimidoyl-2-pyrrolidinyl)methoxy]phenyl]-3-(5-amidinobenzo[b]thien-2-yl)propionicacid;

3-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-4-(5-amidinobenzo[b]thien-2-yl)butyricacid;

2-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-amidino-1-ethyl-2-indolyl)propionicacid;

2-[4-[((3R)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-amidino-1-ethyl-2-indolyl)propionicacid; and

2-[4-[(1-acetimidoyl-4-piperidinyl)oxy]phenyl]-3-(6-amidino-1-ethyl-2-indolyl)propionicacid.

Basically, the compound of formula (1) of the present invention can beproduced for example in accordance with the following reaction formulae.Namely, the nitrile form of the formula (3) is reacted with an alcohol(R⁵ OH) in the presence of a hydrogen halide. The resulting imidate form(4) is reacted with ammonia to obtain an aromatic amidine derivative(1a). ##STR6##

In the above formulae, R¹, R², R³, R⁴, n, A, X, Y

and ##STR7## are the same as described above, and R⁵ is a lower alkylgroup.

The above reaction sequence is described in detail. Reaction of thenitrile form (3) with an alcohol (R⁵ OH) may be effected for example byallowing the nitrile form (3) to react with the equimolar or an excessamount of an alcohol (R⁵ OH) having 1 to 6 carbon atoms, such asmethanol, ethanol, propanol or the like, in the presence of a hydrogenhalide such as hydrogen chloride, hydrogen bromide or the like. Ifnecessary, a solvent may be used which is selected for example fromaliphatic ethers such as diethyl ether and the like, halogenatedhydrocarbons such as chloroform, dichloromethane and the like, aproticsolvents such as benzene and the like, and mixtures thereof. In general,the reaction is carried at a temperature of from -20° C. to 60° C. for aperiod of 3 to 220 hours. Preferably, the reaction may be carried out ata temperature of from -8° C. to 30° C. for a period of 10 to 96 hours inthe presence of an excess amount of methanol or ethanol using ahalogenated hydrocarbon solvent such as chloroform or dichloromethane.

Reaction of the thus obtained imidate form (4) with ammonia may beeffected by allowing the imidate form (4) to react with ammonia in asolvent or a mixed solvent system which is selected for example fromalcohols having 1 to 4 carbon atoms, such as ethanol, propanol and thelike, aliphatic ethers such as diethyl ether and the like, halogenatedhydrocarbons such as chloroform and the like, aprotic solvents such asbenzene and the like, and N,N'-dimethylformamide and dimethylsulfoxide.The reaction may be carried out at a temperature of from -10° C. to 140°C. for a period of 0.5 to 200 hours, preferably at a temperature of from-8° C. to 30° C. for a period of 10 to 96 hours in ethanol.

When the nitrile form (3) to be used as a starting material has acarboxyl group or an alkoxycarbonyl group, the carboxyl oralkoxycarbonyl group is esterified by a formation reaction of an imidateor exposed to ester interchange with the alcohol (R⁵ OH) to be used. Asa consequence, since a carboxyl group in a compound (1a) obtained bythis reaction is esterified, it is necessary to subject the compound(1a) to hydrolysis when an aromatic amidine derivative having a freecarboxyl group is produced.

The hydrolysis reaction may be effected by treating the compound (1a) inan aqueous solution of an inorganic acid Such as hydrochloric acid,sulfuric acid or the like or an organic acid such as tosyl acid or thelike, at a temperature of from -10° C. to a reflux temperature,preferably from -5° C. to a reflux temperature, for a period of from 0.5to 550 hours, preferably from 0.5 to 350 hours.

When the compound (1a) contains a group which is susceptible tohydrolysis by a strong acid, it is preferable to protect the amidinogroup with a protective group such as tert-butoxycarbonyl or the likeprior to the hydrolysis reaction, thereafter carrying out esterhydrolysis under basic conditions and subsequent deprotection.Protection of the amidino group may be effected by allowing the compound(1a) to react with 2-(tert-butoxycarbonyloxyimino)-2-phenylacetonitrilein water, methanol, ethanol, tetrahydrofuran, dioxane, acetone or amixture thereof, in the presence of a base such as1,8-diazabicyclo[5.4.0]-7-undecene or the like. The reaction may becarried out at a temperature of from 0° to 50° C., preferably from 5° to30° C., for a period of from 0.5 to 48 hours, preferably from 1 to 24hours.

Ester hydrolysis of the thus protected compound and subsequentdeprotection may be effected by treatment of the protected compound withan aqueous solution of sodium hydroxide or potassium hydroxide and thenwith hydrochloric acid, in water or a water-containing solvent such asof ethanol, methanol, tetrahydrofuran, dioxane or the like. The esterhydrolysis reaction may be carried out at a temperature of from 0° to50° C., preferably from 5° to 30° C., for a period of from 0.5 to 48hours, preferably from 1 to 24 hours. The deprotection reaction may becarried out at a temperature of from 0° to 60° C., preferably at 25° C.,for a period of from 0.5 to 24 hours, preferably from 1 to 6 hours.

When two alkoxycarbonyl groups are linked to one carbon atom in group Aof the compound (1a), hydrolysis and decarboxylation may be carried outat the same time in accordance with the following reaction sequence:##STR8##

In the above formulae, each of 1 and m is 0 or 1, while R¹, R², R³, R⁴,R⁵, n, X, Y and ##STR9## are the same as described above.

This reaction may be carried out in an aqueous solution of an inorganicacid such as hydrochloric acid, sulfuric acid or the like or an organicacid such as tosyl acid or the like, at a temperature of from -20° C. toa reflux temperature, preferably from -5° C. to a reflux temperature,for a period of from 0.5 to 550 hours, preferably from 0.5 to 350 hours.

When a compound (1e) having an imidoyl group in its group Y is producedas a compound of formula (1) of the present invention, it may beobtained by allowing a compound (1d) having a primary or secondary aminogroup in its group Y to react with an imidate compound (5) in accordancewith the following reaction sequence: ##STR10##

In the above formulae, Y¹ is a variety of the aforementioned Y groupshaving a primary or secondary amino group as a substituent, Y² isanother variety of the aforementioned Y groups having an imidyl group asa substituent, each of R⁶ and R⁷ is a hydrogen atom, a lower alkyl groupor a phenyl group and R⁸ is a lower alkyl group or a benzyl group, whileR¹, R², R³, R⁴, n, A, X and ##STR11## are the same as described above.

This reaction may be effected for example by allowing compound (1d) toreact with the equimolar or an excess amount of imidate compound (5) inthe presence of a base such as triethylamine, sodium hydroxide,potassium hydroxide or the like, in water or a solvent or a mixedsolvent system which is selected for example from alcohols having 1 to 4carbon atoms such as ethanol, propanol and the like, aliphatic etherssuch as diethyl ether and the like, halogenated hydrocarbons such aschloroform and the like, and N,N'-dimethylformamide anddimethylsulfoxide. The reaction may be carried out at a temperature offrom -20° C. to 70° C. for a period of from 1 minute to 168 hours,preferably at a temperature of from -10° C. to 40° C. for a period offrom 1 minute to 72 hours.

When the imidoyl form (1e) has a alkoxycarbonyl group, thealkoxycarbonyl group may be hydrolyzed to a carboxylic group.

The hydrolysis reaction may be effected by treating the compound (1e) inan aqueous solution of an inorganic acid such as hydrochloric acid,sulfuric acid or the like or an organic acid such as tosyl acid or thelike, at a temperature of from -10° C. to a reflux temperature,preferably from -5° C. to a reflux temperature, for a period of from 0.5to 550 hours, preferably from 0.5 to 350 hours.

According to the present invention, when a material compound has asubstituent such as a carboxyl group, an amino group or the like, it ispreferable to protect such a functional group prior to necessaryreactions, and thereafter detaching the protective group. On the otherhand, the formation reaction of an amidine, the formation reaction of animidate and the like may be carried without protecting such a functionalgroup. In this instance, protection of the primary or secondary aminogroup may be effected by use of a protective group such astert-butoxycarbonyl, benzyloxycarbonyl, p-nitrobenzyloxycarbonyl,triphenylmethyl or the like.

In addition, an alkoxycarbonyl-substituted compound may be obtained forexample in accordance with the following reaction sequence, by carryingout ester hydrolysis after the formation reaction of an amidine or animidate, followed, if necessary, by re-esterification: ##STR12##

In the above formulae, R⁹ is a hydrogen atom or a lower alkyl group, R¹⁰is a lower alkyl group and p is an integer of 1 or 2, while R⁵, n, X, Yand ##STR13## are the same as described above.

That is, a nitrile compound represented by formula (6) is allowed toreact with an alcohol (R⁵ OH) in the presence of a hydrogen halide, andthe resulting imidate.ester compound is hydrolyzed by a base treatmentto obtain an imidate-carboxylic acid derivative (7) which issubsequently reacted with ammonia to obtain an amidino group-substitutedaromatic compound (1f). By subjecting compound (1f) to esterification, acompound (1g) is produced.

Reaction of the nitrile compound (6) with an alcohol (R⁵ OH) may beeffected for example by allowing the nitrile compound (6) to react withthe equimolar or an excess amount of an alcohol (R⁵ OH) having 1 to 6carbon atoms, such as methanol, ethanol, propanol or the like, in thepresence of a hydrogen halide such as hydrogen chloride, hydrogenbromide or the like. If necessary, a solvent or a solvent mixture may beused which is selected, for example, from aliphatic ethers such asdiethyl ether and the like, halogenated hydrocarbons such as chloroform,dichloromethane and the like and aprotic solvents such as benzene. Thereaction may be carried out at a temperature of from -10° C. to 60° C.for a period of from 3 to 120 hours. Preferably, it may be effected at atemperature of from -8° C. to 30° C. for a period of from 10 to 96 hoursin a halogenated hydrocarbon solvent such as chloroform ordichloromethane in the presence of an excess amount of methanol orethanol. After concentrating and drying the resulting reaction mixture,the remaining solid material is treated with a strong alkali solution toeffect neutralization and ester hydrolysis, thereby obtaining theimidate.carboxylic acid derivative represented by formula (7). Thereaction may be carried out generally at a temperature of from -10° C.to 60° C. for a period of from 0.2 to 5 hours, preferably at atemperature of from 0° to 25° C. for a period of from 0.5 to 2 hours, inan aqueous solution of sodium hydroxide or potassium hydroxide.

Reaction of the thus obtained imidate-carboxylic acid derivative (7)with ammonia may be effected for example by allowing the derivative (7)to react with ammonium chloride, ammonia or a mixture thereof, in asolvent or a mixed solvent system which is selected for example fromalcohols having 1 to 4 carbon atoms such as ethanol, propanol and thelike, aliphatic ethers such as diethyl ether and the like, halogenatedhydrocarbons such as chloroform and the like, aprotic solvents such asbenzene and the like and N,N-dimethylformamide and dimethylsulfoxide.The reaction may be carried out generally at a temperature of from -10°C. to 140° C. for a period of from 0.5 to 200 hours, preferably at atemperature of from -8° C. to 30° C. for a period of from 10 to 96hours, in ethanol.

Esterification of the amidino compound represented by formula (1f) maybe effected, for example, by allowing compound (1f) to react with athionyl halide such as thionyl chloride, thionyl bromide or the like inan alcohol having 1 to 4 carbon atoms such as ethanol, propanol or thelike. The reaction may be carried out generally at a temperature of from0° C. to a reflux temperature for a period of from 10 minutes to 36hours, preferably at a temperature of from 10° to 60° C. for a period offrom 10 minutes to 24 hours.

Crystallization of a compound of formula (1) of the present inventionmay be effected for example by treating the reaction-completed solutionwith a strongly basic (OH) type ion exchange resin, or with sodiumhydroxide, potassium hydroxide or the like, to adjust the number ofadded salts, preferably to 1. The resulting solution is treated at atemperature of from -10° C. to 30° C., preferably from 0° to 25° C., inwater or a solvent such as methanol, ethanol, isopropanol, acetone orthe like or a mixture thereof, preferably in a water/ethanol mixturesystem.

The thus obtained aromatic amidine derivative of formula (1) or a saltthereof has a specific and excellent capacity to inhibit FXa and isuseful as an anticoagulant agent, as well as a preventive agent and atherapeutic agent for thrombosis and embolism. Since a compound offormula (1) can exhibit its effect even when administered orally, it canbe applied to both oral and parenteral administrations. The compound ofthe present invention may be administered by optionally changing itsdose depending on symptoms, age, weight and the like of each patient. Inthe case of oral administration, the compound may be administeredgenerally in a dose of from 5 to 1,000 mg/day/adult, preferably from 10to 500 mg/day/adult. Examples of dosage forms include tablets, capsules,powders, granules and the like which can be prepared in the usual wayusing generally used additives such as fillers, lubricants, binders andthe like. In the case of parenteral administration, the compound may beadministered by subcutaneous injection, intravenous injection orintravenous drip infusion in a dose of from 0.1 to 100 mg/day/adult,preferably from 0.5 to 30 mg/day/adult.

Since the compound of the present invention shows a high anticoagulantfunction based on its excellent FXa inhibition activity, it does notreact with platelets and can be applied to various diseases caused bythrombosis and embolism, such as cerebral infarction, cerebralthrombosis, cerebral embolism, transient cerebral ischemic attack (TIA),myocardial infarction, unstable angina, pulmonary infarction, pulmonaryembolism, Berger disease, deep venous thrombosis, disseminatedintravascular coagulation syndrome, thrombus formation after artificialblood vessel operation, artificial valve replacement, percutaneoustransluminal colonary angioplasty (PTCA), or percutaneous transluminalcoronary recanalization (PTCR), obstruction after recirculation ofblood, thrombus formation during extracorporeal circulation and thelike.

The following reference, inventive and test examples are provided toillustrate further the present invention. It is to be understood,however, that the examples are for the purpose of illustration only andare not intended as a definition of the limits of the invention.

REFERENCE EXAMPLE 1 Preparation of(5-cyano-3-methyl-2-benzofuranyl)methyltriphenylphosphonium chloride

a) 13.31 g of 2-acetyl-4-bromophenol, 11.0 g of ethyl bromoacetate and9.7 g of anhydrous potassium carbonate were refluxed under heating in 70ml of acetone for 2 hours. Insoluble materials were removed byfiltration, and the resulting filtrate was concentrated and dried. Theresidue thus obtained was dissolved in chloroform, washed with water,and then dried to remove the solvent. The thus treated residue waswashed with a mixed solvent system of ethanol and n-hexane to isolateinsoluble crystals by filtration. In this way, 16.82 g of ethyl(2-acetyl-4-bromophenyl)oxyacetate was obtained in the form of colorlessplate crystals.

mp: 66°-68° C.

b) 16.8 g of ethyl (2-acetyl-4-bromophenyl)oxyacetate obtained in theabove step a) was dissolved in 100 ml of anhydrous ethanol to which 1.2g of metallic sodium has been dissolved in advance, and the resultingsolution was stirred at room temperature for 1.5 hours. The reactionsolution was poured into water and extracted with ethyl acetate, and theresulting organic layer was washed with water, and then dried. Afterdistilling off the solvent, precipitated crystals were collected byfiltration and washed with ethanol to obtain 5.3 g of ethyl5-bromo-3-methyl-2-benzofurancarboxylate in the form of colorless fineneedle crystals.

mp: 96°-97° C.

¹ H-NMR (CDCl₃) δ:1.44 (3H, t, J=8Hz), 2.54 (3H, s), 4.45 (2H, q,J=8Hz), 7.43 (2H), 7.73 (1H, s)

c) In a stream of nitrogen, 4.9 g of ethyl5-bromo-3-methyl-2-benzofurancarboxylate obtained in the above step b),2.0 g of cuprous cyanide and a catalytically effective amount of coppersulfate were stirred in 40 ml of N-methyl-2-pyrrolidone for 6 hours at200° C. After cooling, the reaction solution was poured into water toremove insoluble materials by filtration. The resulting filtrate wasextracted with ethyl acetate, and the organic layer was washed withwater, and then concentrated and dried to collect precipitated crystals.In this way, 3.16 g of ethyl 5-cyano-3-methyl-2-benzofurancarboxylate inthe form of light brown crystals.

mp: 156°-158° C.

¹ H-NMR (CDCl₃) δ:1.45 (3H, t, J=8Hz), 2.60 (3H, s), 4.45 (2H, q,J=8Hz), 7.67 (2H), 7.99 (1H, s)

d) 3.1 g of ethyl 5-cyano-3-methyl-2-benzofurancarboxylate obtained inthe above step c) was dissolved in 60 ml of tetrahydrofuran. To thiswere added, with ice cooling, 2.1 g of calcium iodide (4H₂ O), 0.63 g ofsodium borohydride and a catalytically effective amount of sodiumbicarbonate. The resulting mixture was stirred at room temperature for18 hours, followed by further addition of 2.1 g of calcium iodide (4H₂O) and 0.63 g of sodium borohydride and by additional stirring at roomtemperature for 18 hours.

The resulting reaction solution was diluted with ethyl acetate, washedwith water and then dried to remove the solvent. The residue thusobtained was subjected to silica gel column chromatography usingchloroform as an eluant. In this way, 1.96 g of purified2-hydroxymethyl-3-methyl-5-benzofurancarbonitrile was obtained.

¹ H-NMR (CDCl₃) δ:1.8 (1H, br), 2.28 (3H, s), 4.78 (2H, s), 7.52 (2H),7.82 (1H, s)

e) 1.92 g of 2-hydroxymethyl-3-methyl-5-benzofurancarbonitrile obtainedin the above step d) was added to 50 ml of diethyl ether, followed bythe addition of 3 drops of pyridine and 1.65 ml of thionyl chlorideduring ice cooling, and the resulting mixture was stirred at roomtemperature for 4.5 hours. The reaction solution was poured into icewater and extracted with chloroform, and the resulting organic layer waswashed with water, saturated sodium bicarbonate aqueous solution andwater in that order, followed by concentration and drying. In this way,1.68 g of 2-chloromethyl-3-methyl-5-benzofurancarbonitrile was obtained.

f) 1.68 g of 2-chloromethyl-3-methyl-5-benzofurancarbonitrile obtainedin the above step e) and 3 g of triphenylphosphine were refluxed underheating in xylene for 5 hours. After cooling, precipitated crystals werecollected by filtration to obtain 3.63 g of the title compound.

mp: >270° C.

¹ H-NMR (CDCl₃) δ: 2.0 (1.5H, s), 2.04 (1.5H, s), 6.09 (2H, d, J=16Hz),7.7 (18H, m)

REFERENCE EXAMPLE 2 Preparation of(5-cyano-3-benzofuranyl)methyltriphenylphosphonium bromide

a) 12.15 g of ethyl 5-cyano-3-methyl-2-benzofurancarboxylate obtained inthe step c) of Reference Example 1 was dissolved in 60 ml of ethanol,followed by the addition of 5 g of sodium hydroxide and 100 ml of water,and the resulting mixture was stirred at 30° to 40° C. for 2 hours.After ice cooling, the resulting reaction solution was adjusted to pH 2with dilute HCl solution, and crystals thus precipitated were collectedby filtration and dried. In this way, 10.6 g of5-cyano-3-methyl-2-benzofurancarboxylic acid was obtained in the form ofcolorless prism crystals.

mp: (sublimation at 275°-285° C.)

¹ H-NMR (CDCl₃) δ:2.54 (3H, s), 7.88 (2H), 8.44 (1H)

b) 10.64 g of 5-cyano-3-methyl-2-benzofurancarboxylic acid obtained inthe above step a) and 2.5 g of copper powder were added to 65 ml ofquinoline, and the mixture was stirred at 210° C. for 30 minutes. Afteradding lice water and adjusting to pH 1 with HCl, the reaction mixturethus treated was extracted with chloroform, and the resulting organiclayer was dried under a reduced pressure. The residue thus obtained wassubjected to silica gel column chromatography using toluene as aneluant. In this way, 6.89 g of purified colorless3-methyl-5-benzofurancarbonitrile was obtained.

mp: 73° C.

¹ H-NMR (CDCl₃) δ: 2.26 (3H, d, J=1.5Hz), 7.53 (3H), 7.85 (1H, s)

c) 7.28 g of 3-methyl-5-benzofurancarbonitrile obtained in the abovestep b) was dissolved in 50 ml of carbon tetrachloride and subjected toreflux under light irradiation condition. To the resulting reactionsolution was gradually added a mixture consisting of 8.25 g ofN-bromosuccinimide and 160 mg of 2,2-azobis-iso-butylonitrile. Afterrefluxing under heating for 3 hours, precipitated materials were removedby filtration, and the resulting filtrate was dried. The thus driedresidue was subjected to purification by silica gel columnchromatography using toluene as an eluant, thereby obtaining 8.65 g of amixture (2:5) of the starting material 3-methyl-5benzofurancarbonitrileand 3-bromomethyl-5-benzofurancarbonitrile. 8.65 g of the thus obtainedcrude bromomethyl compound was dissolved in xylene, 10 g oftriphenylphosphine was added to the resulting solution, and the thusprepared mixture was heated for 20 minutes. After cooling, theprecipitate thus formed was collected by filtration to obtain 14.73 g ofthe title compound in the form of colorless crystals.

mp: >290° C.

¹ H-NMR (CDCl₃) δ:5.88 (2H, d, J=16Hz), 7.0-8.0 (19H, m)

REFERENCE EXAMPLE 3 Preparation of(5-cyano-7-methoxy-2-benzofuranyl)methyltriphenylphosphonium chloride

a) 10.0 g of 5-bromo-2-hydroxy-3-methoxybenzaldehyde was dissolved in 39ml of N,N-dimethylformamide, and the resulting solution was mixed with11.9 g of anhydrous potassium carbonate and stirred at room temperature.5.0 g of chloroacetone was added dropwise to the above reaction solutionat the same temperature, followed by additional 1 hour of stirring at anelevated temperature of 80° C. The resulting reaction solution wasdiluted with ethyl acetate and adjusted to pH 2 with concentratedhydrochloric acid, and the resulting organic layer was collected. Theorganic layer was dried to distill off the solvent, and the resultingresidue was purified by silica gel column chromatography, therebyobtaining 4.0 g of 2-acetyl-5-bromo-7-methoxybenzofuran.

mp: 107°-109° C.

¹ H-NMR (CDCl₃) δ:2.62 (3H, s), 3.83 (3H, s), 7.02 (1H), 7.39 (2H)

b) To 107.6 ml of 5N sodium hydroxide aqueous solution was addeddropwise 26.8 g of bromine at a temperature of -5° C. or below. To thiswas further added dropwise and slowly a 100 ml dioxane solutioncontaining 15.0 g of 2-acetyl-5-bromo-7-methoxybenzofuran obtained inthe above step a). After completion of the dropwise addition, thetemperature of the resulting reaction solution was increased graduallyto 60° C. and then stirred for 30 minutes. After cooling, the resultingreaction solution was adjusted to pH 2 with concentrated hydrochloricacid and then extracted with ethyl acetate. The resulting organic layerwas concentrated to dryness, and crystals thus precipitated werecollected by filtration to obtain5-bromo-7-methoxy-2-benzofurancarboxylic acid. The thus obtainedcrystals were suspended in 200 ml of ethanol, and 10 ml of thionylchloride was added dropwise to the suspension with stirring at roomtemperature. The resulting reaction solution was refluxed under heatingfor 2 hours. After cooling, the thus treated reaction solution wasneutralized with saturated sodium bicarbonate aqueous solution, and thenmixed with water to collect precipitated crystals by filtration. Thecrystals thus collected were purified by silica gel columnchromatography using chloroform as an eluant to obtain 11.33 g of ethyl5-bromo-7-methoxy-2-benzofurancarboxylate.

¹ H-NMR (CDCl₃) δ:1.41 (3H, t, J=7.0Hz), 4.00 (3H, s), 4.43 (2H, q,J=7Hz), 7.02 (1H, d), 7.39 (1H, d), 7.42 (1H, s)

c) a mixture consisting of 2.0 g of ethyl5-bromo-7-methoxy-2-benzofurancarboxylate obtained in the above step b),1.26 g of cuprous cyanide, 100 ml of N-methyl-2-pyrrolidone and acatalytically effective amount of copper sulfate was stirred at 180° to190° C. for 2 hours in a stream of argon. After cooling, a toluene/ethylacetate mixture (1:1) and water were added to the reaction solution toremove insoluble materials, and the resulting organic layer was washedwith water, and then dried. After distilling off the solvent,precipitated crystals were collected by filtration and washed withethanol to obtain 1.2 g of ethyl5-cyano-7-methoxy-2-benzofurancarboxylate.

¹ H-NMR (CDCl₃) δ:1.43 (3H, t, J=7.0Hz), 4.06 (3H, s), 4.46 (2H, t,J=7.0Hz), 7.10 (1H, d, J=1.0Hz), 7.53 (1H, s), 7.64 (1H, d)

d) 8.55 g of ethyl 5-cyano-7-methoxy-2-benzofurancarboxylate obtained inthe above step c) was dissolved in 250 ml of tetrahydrofuran. Duringcooling on an ice bath, the resulting solution was mixed with 13.74 g ofcalcium iodide (4H₂ O), 2.12 g of sodium borohydride and a catalyticallyeffective amount of sodium bicarbonate, and the resulting mixture wasstirred at room temperature for 1.5 hours, followed by further additionof 13.74 g of calcium iodide (4H₂ O) and 2.12 g of sodium borohydrideand additional stirring at room temperature for 1 hour. During coolingon an ice bath, the resulting reaction solution was adjusted to pH 2with concentrated hydrochloric acid, and the solvent was removed bydistillation. The resulting residue was extracted with chloroform,washed with water and then dried to distill off the solvent. The thustreated residue was purified by silica gel column chromatography using amixture of chloroform and ethanol as an eluant, thereby obtaining 1.96 gof 2-hydroxymethyl-7-methoxy-5-benzofurancarbonitrile.

mp: 149°-150° C.

¹ H-NMR (CDCl₃) δ: 2.17 (1H, t, J=6.1Hz), 4.02 (3H, s), 4.80 (2H, d,J=6.1Hz), 6.71 (1H, s), 6.99 (1H, d, J=1.3Hz), 7.50 (1H, d, J=1.3Hz)

e) 5.0 g of 2-hydroxymethyl-7-methoxy-5-benzofurancarbonitrile obtainedin the above step d) was dissolved in 100 ml of diethyl ether, followedby the addition of a few drops of pyridine. With cooling on an ice bathand with stirring, 5.86 g of thionyl chloride was added dropwise to theabove solution. After completion of the dropwise addition, temperatureof the resulting solution was increased gradually to room temperature,and the stirring was continued for additional 1 hour at the roomtemperature. During cooling on an ice bath, water was added to theresulting reaction solution, and the thus formed organic layer wascollected, washed with water and then dried to remove the solvent,thereby obtaining 2-chloromethyl-7-methoxy-5-benzofurancarbonitrile. Thechloromethyl compound thus obtained and 9.67 g of triphenylphosphinewere refluxed under heating for 18 hours in 50 ml of xylene. Aftercooling, crystals thus precipitated were collected by filtration toobtain 10.54 g of the title compound.

¹ H-NMR (DMSO-d₆) δ:3.89 (3H, s), 5.6-6.0 (2H, br)

REFERENCE EXAMPLE 4 Preparation of(5-cyanobenzo[b]thien-2-yl)methyltriphenylphosphonium chloride

a) 8.13 g of 5-bromosalicylaldehyde was dissolved in 100 ml of acetone,followed by the addition of 6.7 g of anhydrous potassium carbonate. Withstirring at room temperature, 5.0 g of N,N-dimethylthiocarbamoylchloride was added to the above solution, and the stirring was continuedfor two hours. The resulting reaction solution was poured into icewater, and crystals thus precipitated were collected by filtration, anddried to obtain 9.2 g of5-bromo-2-[(N,N-dimethylthiocarbamoyl)oxy]benzaldehyde.

mp: 141°-143° C.

IR (KBr): 1690, 1596, 1546, 1470, 1396 cm⁻¹

¹ H-NMR (CDCl₃) δ: 3.42 (3H, s), 3.47 (3H, s), 7.03 (1H, d, J=8.3Hz),7.72 (1H, dd, J=8.3 and 2.2Hz), 8.01 (1H, d, J=2.2Hz)

b) 9.0 g of 5-bromo-2-[(N,N-dimethylthiocarbamoyl)oxy]benzaldehydeobtained in the above step a) was melted by heating it for 10 minutes onan oil bath of 210° to 220° C. The resulting product was dissolved in a1 ml of toluene, followed by the addition of 6 ml of methanol. Crystalsthus precipitated were collected by filtration to obtain 4.0 g of crude5-bromo-2-[(N,N-dimethylcarbamoyl)thio]benzaldehyde.

mp: 118°-120° C.

IR (KBr): 1677, 1365, 1185 cm⁻¹

¹ H-NMR (CDCl₃) δ:3.09 (6H, s), 7.31 (1H, d, J=9.6Hz), 7.70 (1H, dd,J=9.6 and 1.8Hz), 8.14 (1H, d, J=1.8Hz), 10.25 (1H, s)

c) 21.0 g of 5-bromo-2-[(N,N-dimethylcarbamoyl)thio]benzaldehyde wasdissolved in 50 ml of methyl orthoformate. The resulting solution wasmixed with 1.0 g of p-toluenesulfonate, and refluxed under heating for50 minutes. After cooling, the resulting reaction solution was pouredinto saturated sodium bicarbonate solution and extracted with benzene.The resulting organic layer was dried to remove the solvent. The residuethus obtained was dissolved in 100 ml of methanol, followed by adding 37ml of 2N sodium hydroxide and by refluxing under heating for 1 hour in astream of nitrogen. After cooling, the resulting reaction solution wasadjusted to pH 1 with concentrated hydrochloric acid, extracted withbenzene, and then dried to remove the solvent. The residue thus obtainedwas dissolved in 20 ml of acetone and added dropwise, at roomtemperature, to a stirred mixture consisting of 6.74 g of chloroacetone,22.1 g of anhydrous potassium carbonate and 150 ml of acetone. After 30minutes of stirring, the resulting reaction mixture was refluxed underheating for 30 minutes. After cooling, insoluble materials were removedby filtration, and the resulting filtrate was concentrated to dryness.The thus obtained residue was purified by silica gel columnchromatography using toluene as an elution solvent and the resultingproduct was recrystallized from ethanol to obtain 7.5 g of2-acetyl-5-bromobenzo[b]thiophene.

mp: 120°-121° C.

IR (KBr): 1668, 1512, 1326, 1266 cm⁻¹

¹ H-NMR (CDCl₃) δ:2.67 (3H, s), 7.54 (1H, dd, J=8.8 and 1.8Hz), 7.75(1H, d, J=8.8Hz), 7.85 (1H, s), 8.03 (1H, d, J=1.8Hz)

d) With stirring, 5.4 ml of bromine was added dropwise to 5N sodiumhydroxide aqueous solution which has been cooled to -5° C. to 0° C. Tothis was added dropwise, at a temperature of -5° C. or below, a 50 mldioxane solution of 2-acetyl-5-bromobenzo[b]thiophene obtained in theabove step c). The resulting mixture was stirred for 30 minutes at roomtemperature and then for 30 minutes at 50° C. With ice cooling, theresulting reaction solution was adjusted to pH 2 with concentratedhydrochloric acid, and crystals thus precipitated were collected byfiltration and washed with water. The crystals thus obtained weredissolved in ethyl acetate, and the solution was dried and concentrated.Crystals thus precipitated were collected by filtration and washed withtoluene to obtain 6.6 g of 5-bromobenzo[b]-thiophene-2-carboxylic acid.

mp: 238°-241° C.

IR (KBr): 1671, 1554, 1518, 1443 cm⁻¹

¹ H-NMR (CDCl₃) δ:7.57 (1H, dd, J=8.6 and 1.8Hz), 7.82 (1H, d, J=8.6Hz),8.00 (1H, s), 8.07 (1H, d, J=1.8Hz)

e) 6.4 g of 5-bromobenzo[b]thiophene-2-carboxylic acid obtained in theabove step d) was suspended in 250 ml of ethanol. With cooling on an icebath and with stirring, 4.45 g of thionyl chloride was added dropwise tothe suspension prepared above, followed by refluxing under heating for 1hour. With ice cooling, 8.15 g of thionyl chloride was further addeddropwise to the resulting mixture, followed by refluxing under heating 2hours. The resulting reaction solution was concentrated and adjusted topH 9 with saturated sodium bicarbonate aqueous solution. Crystals thusprecipitated were collected by filtration and dried to obtain 7.0 g ofethyl 5-bromobenzo[b]thiophene-2-carboxylate. A portion of thee thusobtained compound was recrystallized from methanol to obtain needlecrystals.

mp: 94°-95° C.

¹ H-NMR (CDCl₃) δ:1.42 (3H, t, J=7.0Hz), 4.41 (2H, q, J=7.0Hz), 7.54(1H, dd, J=8.8 and 1.8Hz), 7.73 (1H, d, J=8.8Hz), 7.96 (1H, s), 8.01(1H, d)

f) 7.0 g of ethyl 5-bromobenzo[b]thiophene-2-carboxylate obtained in theabove step e) and 5.4 g of cuprous cyanide were suspended in 70 ml ofN-methyl-2-pyrrolidone, and the suspension was stirred for 2 hours withheating at a temperature of 200° C. in a stream of nitrogen. Aftercooling, the reaction mixture was diluted with ethyl acetate, insolublematerials were removed by filtration, and the resulting filtrate waswashed with water and dried. After distilling off the solvent, thecrystals precipitated were collected by filtration and washed withethanol to obtain 5.02 g of ethyl 5-cyanobenzo[b]thiophene-2-carboxylateas crystals.

mp: 138°-139° C.

IR (KBr): 2232, 1728, 1262 cm⁻¹

¹ H-NMR (CDCl₃) δ:1.43 (3H, t, J=7.0Hz), 4.45 (2H, q, J=7.0Hz), 7.70(1H, dd, J=9.0 and 1.8Hz), 8.04 (1H, d, J=9.0Hz), 8.08 (1H), 8.20 (1H)

g) To 150 ml of tetrahydrofuran were added 4.92 g of ethyl5-cyanobenzo[b]thiophene-2-carboxylate obtained in the above step f) andthen 3.33 g of calcium iodide (4H₂ O). With ice cooling and stirring,1.0 g of sodium borohydride and a catalytically effective amount ofsodium bicarbonate were added to the above mixture, and the resultingmixture was stirred at room temperature for 1 hour. After furtheraddition of 3.33 g of calcium iodide (4H₂ O), 1.0 g of sodiumborohydride was added to the mixture which was cooled on an ice bathwith stirring, and the resulting mixture was stirred at roomtemperature. After stirring for 1 hour, 3.33 g of calcium iodide (4H₂ O)was again added to the stirred mixture, followed by further addition of1.0 g of sodium borohydride to the resulting mixture which was cooled onan ice bath with stirring and by subsequent stirring at room temperaturefor 1 hour. The thus obtained reaction solution was diluted with water,extracted with ethyl acetate, and then dried to remove the solvent.Thereafter, the crystals thus precipitated were collected by filtrationand washed with a mixture of benzene and n-hexane to obtain 4.0 g of2-hydroxymethylbenzo[b]thiophene-5-carbonitrile.

mp: 78°-79° C.

IR (KBr): 3496, 2236, 1026 cm⁻¹

¹ H-NMR (CDCl₃) δ: 4.97 (2H, s) 7.26 (1H), 7.51 (1H, dd, J =8.3 and1.8Hz), 7.90 (1H, d, J=8.3Hz), 8.03 (1H)

h) 4.0 g of 2-hydroxymethylbenzo[b]thiophene-5-carbonitrile obtained inthe above step g) was dissolved in 100 ml of diethyl ether, followed bythe addition of 0.1 ml of pyridine. A 5 ml diethyl ether solution of 5.5g of thionyl chloride was added to the above solution under ice coolingand stirring, and the resulting mixture was stirred at room temperaturefor 2 hours. The resulting reaction solution was poured into ice waterand extracted with benzene. The resulting organic layer was washed withsaturated sodium bicarbonate aqueous solution and concentrated todryness. The residue thus obtained was dissolved in 100 ml of xylene,and the solution was mixed with 7.2 g of triphenylphosphine, andrefluxed under heating for 10 hours. Thereafter, the thus precipitatedcrystals were collected by filtration to obtain 6.3 g of the titlecompound.

mp: 271°-274° C. (decomposition)

¹ H-NMR (CDCl₃) δ:6.70 (2H, d, J=15.1Hz), 7.30-8.10 (19H, m)

REFERENCE EXAMPLE 5 Preparation of(7-cyano-2-naphthyl)methyltriphenylphosphonium bromide

a) 11.0 g of 7-methyl-2-naphthalenecarboxylic acid obtained inaccordance with the procedure disclosed in Australian Journal ofChemistry (vol.18, pp. 1351-1364, 1965) was mixed with 70 ml of thionylchloride and refluxed under heating for 4 hours. The resulting reactionsolution was concentrated to dryness. To the residue thus obtained wasadded 300 ml of concentrated aqueous ammonia under cooling. The mixturewas stirred at room temperature for 3 hours, and then extracted withethyl acetate. The resulting organic layer was washed with water andthen with saturated sodium chloride aqueous solution, followed by dryingand removing the solvent. In this way, 8.5 g of7-methyl-2-naphthalenecarboxamide was obtained in the form of colorlessneedle crystals.

mp: 210° to 212° C.

¹ H-NMR (DMSO-d₆) δ: 2.50 (3H, s), 7.4-8.5 (6H, m)

b) 8.0 g of 7-methyl-2-naphthalenecarboxamide obtained in the above stepa) was suspended in 100 ml of tetrahydrofuran, to which was furtheradded, at room temperature, a 100 ml carbon tetrachloride solutioncontaining 22.66 g of triphenylphosphine. The resulting mixture wasstirred at room temperature for 30 minutes and then at 60° C. for 40hours. After cooling to room temperature, insoluble materials wereremoved by filtration, and the resulting filtrate was concentrated undera reduced pressure. 28.35 g of the resulting residue was applied tosilica gel column chromatography, and eluted with a mixed solvent systemof n-hexane and ethyl acetate, thereby obtaining 5.73 g of7-methyl-2-naphthalenecarbonitrile in the form of colorless crystals.

mp: 134°-136° C.

¹ H-NMR (CDCl₃) δ:2.54 (3H, s)i, 7.4-8.2 (6H, m)

c) 5.7 g of 7-methyl-2-naphthalenecarbonitrile obtained in the abovestep b) was suspended in 100 ml of carbon tetrachloride. To this wereadded 6.37 g of N-bromosuccinimide and 30 mg of2,2-azobis-iso-butylonitrile. After refluxing under heating for 2 hours,the resulting reaction solution was diluted with dichloromethane, washedwith water and then with saturated sodium chloride aqueous solution,followed by drying. By distilling off the solvent, 8.34 g of7-bromomethyl-2-naphthalenecarbonitrile was obtained in the form oflight yellow needle crystals.

mp: 110°-116° C.

¹ H-NMR (CDCl₃) δ:4.65 (2H, s), 7.55-8.28 (6H, m)

d) 8.34 g of 7-bromomethyl-2-naphthalenecarbonitrile obtained in theabove step c) was dissolved in 200 ml of xylene, the solution was mixedwith 11.6 g of triphenylphosphine and the mixture was refluxed underheating for 16 hours. Diethyl ether was added to the resulting reactionsolution, and crystals thus precipitated were collected by filtrationand dried, thereby obtaining 12.10 g of the title compound.

¹ H-NMR (CDCl₃) δ:5.96 (2H, d, J=15.3Hz), 7.1-8.0 (21H, m)

REFERENCE EXAMPLE 6 Preparation of(6-cyano-1-methyl-2-indolyl)methyltriphenylphosphonium bromide

a) 1.5 g of methyl 6-cyano-2-indolecarboxylate obtained in accordancewith the procedure disclosed in Liebigs Annalen der Chemie (1986, pp.438-455) was dissolved in 20 ml of N,N-dimethylformamide. 320 mg of 60%sodium hydride was added to the above solution under ice cooling andstirring, and the resulting mixture was stirred at room temperature for10 minutes. To this was further added 0.47 ml of methyl iodide, followedby stirring at room temperature for 2 hours. Saturated ammonium chlorideaqueous solution was added to the resulting reaction solution, andcrystals thus precipitated were collected by filtration, and washed withmethanol. The thus washed crystals were recrystallized from a mixture ofdichloromethane and methanol to obtain 1.4 g of methyl6-cyano-1-methyl-2-indolecarboxylate.

¹ H-NMR (DMSO-d₆) δ:3.92 (3H, s), 4.10 (3H, s), 7.42 (1H, s), 7.52 (1H,dd), 7.98 (1H, d), 8.38 (1H, br)

b) 5.7 g of methyl 6-cyano-1-methyl-2-indolecarboxylate obtained in theabove step a) was dissolved in 120 ml of tetrahydrofuran. With coolingon an ice bath and with stirring, a catalytically effective amount ofsodium bicarbonate, 5.6 g of calcium iodide and 1.8 g of sodiumborohydride were added to the above solution, and the mixture wasstirred for 5 hours. The resulting reaction solution was mixed with icewater and acetic acid, tetrahydrofuran was distilled off from themixture, and the thus treated reaction solution was extracted with ethylacetate, followed by drying. After distilling off the solvent, theresidue thus obtained was dissolved in 50 ml of dichloromethane. 10 mldichloromethane solution containing 1 ml of phosphorus tribromide wasadded dropwise to the above reaction solution under ice cooling andstirring, and the resulting mixture was stirred at the same temperaturefor 2 hours and then at room temperature for 2 hours. The thus treatedreaction solution was mixed with ice water, washed with sodium carbonateaqueous solution, and then dried. The resulting organic layer wasconcentrated by a factor of about 2 under a reduced pressure, mixed with15 g of triphenylphosphine, and then refluxed under heating for 12hours. Thereafter, the precipitate thus formed was collected byfiltration to obtain 10.5 g of the title compound.

¹ H-NMR (DMSO-d₆) δ:3.33 (3H, s), 5.55 (2H, d), 6.26 (1H, s), 7.20-8.10(18H, m)

The following compounds of Reference Examples 7 and 8 were prepared in amanner similar to the procedure of Reference Example 6.

REFERENCE EXAMPLE 7(6-cyano-1-ethyl-2-indolyl)methyltriphenylphosphonium bromide

¹ H-NMR (DMSO-d₆) δ: 1.01 (3H, t), 3.83 (2H), 5.57 (2H, d), 6.26 (1H,s), 7.39 (1H, d), 7.59 (1H, d), 7.70-8.00 (16H, m)

REFERENCE EXAMPLE 8[1-(2-chloroethyl)-6-cyano-2-indolyl]methyltriphenylphosphonium bromide

¹ H-NMR (DMSO-d₆) δ:3.40-3.80 (2H), 4.30-4.60 (2H), 5.60 (2H, d), 6.25(1H, s), 7.10-8.00 (18H, m)

REFERENCE EXAMPLE 9 Preparation of2-bromomethyl-5-benzothiazolecarbonitrile

a) 28.0 g of 5-bromo-2-methylbenzothiazole was dissolved in 200 ml ofN-methyl-2-pyrrolidone, the solution thus prepared was mixed with 13.8 gof cuprous cyanide and a catalytically effective amount of coppersulfate, and the mixture was stirred for 4 hours with heating at atemperature of 180° to 190° C. in a stream of nitrogen. The resultingreaction solution was poured into water, and insoluble materials thusformed were collected by filtration. The thus collected insolublematerials were mixed with a mixture consisting of 22 ml ofmethylenediamine and 50 ml of water, and the resulting mixture wasstirred thoroughly. After extraction with benzene, the resulting organiclayer was washed with water, and dried to distill of benzene.Thereafter, the residue thus formed was washed with ethanol to obtain10.22 g of 2-methyl-5-benzothiazolecarbonitrile in the form of lightbrown crystals.

mp: 158°-160° C.

¹ H-NMR (CDCl₃) δ:2.90 (3H, s), 7.60 (1H, dd), 7.95 (1H, d), 8.25 (1H,d)

b) 7.46 g of 2-methyl-5-benzothiazolecarbonitrile obtained in the abovestep a) was dissolved in 250 ml of carbon tetrachloride, and thesolution was subjected to reflux under a light irradiation condition. Tothe resulting reaction solution was gradually added a mixture consistingof 7.62 g of N-bromosuccinimide and 150 mg of2,2-azobis-iso-butylonitrile, followed by refluxing under heating for 20hours. After cooling, insoluble materials were removed by filtration,and the solvent was distilled off. Thereafter, the residue thus formedwas purified by silica gel column chromatography using toluene as anelution solvent, thereby obtaining 2.18 g of the title compound in theform of light yellow prism crystals.

mp: 185°-186° C.

¹ H-NMR (CDCl₃) δ:4.83 (2H, s), 7.67 (1H, dd), 8.02 (1H, d), 8.34 (1H,d)

REFERENCE EXAMPLE 10 Preparation of(6-cyano-1,2,3,4-tetrahydro-2-naphthyl)-methyltriphenylphosphoniump-toluene sulfonate

a) 10.0 g of methyl6-hydroxymethyl-5,6,7,8-tetrahydro-2-naphthalenecarboxylate was added to3.82 g of 2,3-dihydropyrane. After further adding 5 drops ofconcentrated sulfuric acid, the resulting mixture was stirred for 1hour. To this were further added 1.00 g of 2,3-dihydropyrane and 3 dropsof concentrated sulfuric acid, followed by stirring for 5 hours. Theresulting reaction mixture was mixed with 100 ml of diethyl ether, andthe mixture was washed with saturated sodium bicarbonate aqueoussolution, water and saturated sodium chloride aqueous solution in thatorder, followed by drying. By distilling off the solvent, 13.72 g ofmethyl6-[(2-tetrahydropyranyl)oxymethyl]-5,6,7,8-tetrahydro-2-naphthalenecarboxylatewas obtained in the form of yellow oil.

¹ H-NMR (CDCl₃) δ: 1.50-3.00 (13H, m), 3.30-4.10 (4H, m), 3.86 (3H, s),4.60 (1H, br), 7.10 (1H, d), 7.80-7.90 (2H, m)

b) 13.72 g of methyl6-[(2-tetrahydropyranyl)oxymethyl]-5,6,7,8-tetrahydro-2-naphthalenecarboxylateobtained in the above step a) was dissolved in 180 ml of methanol. Afterfurther adding a solution containing 2.96 g of sodium hydroxide in 60 mlof water, the resulting mixture was refluxed under heating for 3 hours.After cooling, the resulting reaction solution was concentrated underreduced pressure, mixed with chloroform and water, and then neutralizedwith acetic acid. The resulting organic layer was washed with water andthen with saturated sodium chloride aqueous solution, followed bydrying. After distilling off the solvent, the resulting residue wascrystallized in isopropyl ether to obtain 10.51 g of6-[(2-tetrahydropyranyl)oxymethyl]-5,6,7,8-tetrahydro-2-naphthalenecarboxylicacid.

¹ H-NMR (CDCl₃) δ:1.50-3.00 (13H, m), 3.30-4.00 (4H, m), 4.60 (1H, br),7.16 (1H, d), 7.80-7.90 (2H, m)

c) 12.0 g of6-[(2-tetrahydropyranyl)oxymethyl]-5,6,7,8-tetrahydro-2-naphthalenecarboxylicacid obtained in the above step b) and 4.1 g of triethylamine weredissolved in 100 ml of tetrahydrofuran, and the resulting solution wascooled down to -15° C. To this was added, with stirring, 5.64 g ofchloroformic isobutyl ester. The resulting reaction solution was stirredfor 20 minutes at the same temperature and then poured in 200 ml ofice-cold ethanol containing 14% (w/v) of ammonia. After removinginsoluble materials by filtration, the resulting filtrate was driedunder a reduced pressure. The residue thus obtained was purified bysilica gel column chromatography using a mixed solvent system consistingof n-hexane and ethyl acetate as an elution solvent, and the purifiedproduct was crystallized in isopropyl ether to obtain 7.20 g of6-[(2-tetrahydropyranyl)oxymethyl]-5,6,7,8-tetrahydro-2-naphthalenecarboxamide.

¹ H-NMR (CDCl₃) δ:1.40-3.00 (13H, m), 3.30-4.00 (4H, m), 4.60 (1H, br),6.10 (2H, br), 7.20 (1H, d), 7.50-7.70 (2H, m)

d) 15 0 g of6-[(2-tetrahydropyranyl)oxymethyl]-5,6,7,8-tetrahydro-2-naphthalenecarboxamideobtained in the above step c) was suspended in 60 ml of dioxane. Afteradding 8.35 ml of pyridine, the resulting suspension was cooled down to-8° C. to 0° C.

Then, 7.89 ml of anhydrous trifluoroacetate was added dropwise theretounder stirring. The resulting reaction solution was stirred at -5° C.for 30 minutes and then at room temperature for 2 hours. The thustreated reaction solution was diluted with chloroform, and then washedwith water and saturated sodium chloride aqueous solution in that order.Thereafter, the resulting organic layer was dried and the solvent wasdistilled off to obtain 9.78 g of6-[(2-tetrahydropyranyl)oxymethyl]-5,6,7,8-tetrahydro-2-naphthalenecarbonitrilein the form of oil.

¹ H-NMR (CDCl₃) δ:1.50-3.00 (13H, m), 3.30-4.00 (4H, m), 4.61 (1H, br),7.05-7.50 (3H, m)

e) 9.78 g of6-[(2-tetrahydropyranyl)oxymethyl]-5,6,7,8-tetrahydro-2-naphthalenecarbonitrilewas dissolved in 100 ml of ethanol. After adding 100 mg ofp-toluenesulfonic acid, the resulting mixture was stirred at roomtemperature for 15 hours. The resulting reaction solution wasneutralized with saturated sodium bicarbonate aqueous solution, followedby the removal of the solvent by distillation. The residue thus obtainedwas dissolved in chloroform, and the solution was washed with water andthen with saturated sodium chloride aqueous solution. The resultingorganic layer was dried and the solvent was distilled off. Thereafter,5.26 g of 6-hydroxymethyl-5,6,7,8-tetrahydro-2-naphthalenecarbonitrilewas crystallized from isopropanol in the form of colorless crystals.

mp: 83°-85° C.

¹ H-NMR (CDCl₃) δ:1.30-3.00 (7H, m), 3.64 (2H, d, J=6.0Hz), 7.05-7.50(3H, m)

f) 15.0 g of6-hydroxymethyl-5,6,7,8-tetrahydro-2-naphthalenecarbonitrile obtained inthe above step e) and 30.5 g of p-toluenesulfonyl chloride weredissolved in 150 ml of pyridine, and the solution was stirred at roomtemperature for 15 hours. The resulting reaction solution was pouredinto ice water, and the thus precipitated crystals were collected byfiltration, washed with water and isopropanol in that order, and thendried. In this way, 24.72 g of colorless5,6,7,8-tetrahydro-6-[(p-toluenesulfonyl)oxymethyl]-2-naphthalenecarbonitrilewas obtained.

mp: 100°-102° C.

¹ H-NMR (CDCl₃) δ:1.20-3.80 (7H, m), 2.47 (3H, s), 4.00 (2H, d,J=6.0Hz), 7.10 (1H, d, J=9.0Hz), 7.30-7.50 (4H, m), 7.80 (2H, d)

g) 24.00 g of5,6,7,8-tetrahydro-6-[(p-toluenesulfonyl)oxymethyl]-2-naphthalenecarbonitrileand 18.38 g of triphenylphosphine were mixed, and then heated at atemperature of 130° to 140° C. for 15 hours in a sealed container. Theresulting reaction product was crystallized from an acetone/n-hexanemixture to obtain 23.3 g of the title compound in the form of lightyellow powder.

¹ H-NMR (CDCl₃) δ:1.40-2.90 (7H, m), 2.27 (3H, s), 3.60-3.90 (2H, m),6.80-7.30 (5H, m), 7.40-8.00 (17H, m)

REFERENCE EXAMPLE 11 Preparation of(6-cyano-2-naphthyl)methyltriphenylphosphonium bromide

a) 6.11 g of 6-methyl-2-naphthalenecarbonitrile was dissolved in 100 mlof carbon tetrachloride, and the solution was mixed with 6.63 g ofN-bromosuccinimide and 30 mg of 2,2-azobis-iso-butylonitrile. Afterrefluxing under heating for 4 hours, the resulting reaction solution wasmixed with chloroform, washed with water, and then dried. By distillingoff the solvent, 7.07 g of colorless6-bromomethyl-2-naphthalenecarbonitrile was obtained.

mp: 134°-137° C.

¹ H-NMR (CDCl₃) δ: 4.65 (2H, s), 7.60-7.80 (2H, m), 7.80-8.00 (3H, m),8.22 (1H, s)

b) 2.0 g of 6-bromomethyl-2-naphthalenecarbonitrile obtained in theabove step a) and 2.77 g of triphenylphosphine were dissolved in 50 mlof xylene. After refluxing under heating for 18 hours, precipitatedcrystals were collected by filtration to obtain 3.31 g of the titlecompound.

mp: >270° C.

¹ H-NMR (CDCl₃) δ: 5.93 (2H, d, J=15.2Hz), 7.40-8.00 (21H, m)

REFERENCE EXAMPLE 12 Preparation of (S)-(+)-3-hydroxytetrahydrofuran

0.23 g of p-toluenesulfonic acid was added to 25 g of(S)-(-)-1,2,4-butanetriol, and the mixture was stirred at 100° C. for 5minutes and then at 180° to 200° C. for 10 minutes. The resultingreaction mixture was subjected to distillation to collect a fraction of95°-100° C./30 mmHg, thereby obtaining 16.2 g of the title compound asan oily material.

¹ H-NMR (CDCl₃) δ:1.80-2.20 (2H, m), 3.76 (2H, d), 3.70-4.10 (2H, m),4.40-4.60 (1H, m)

REFERENCE EXAMPLE 13 Preparation of ethyl2-[4-[[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-2-oxoacetate

To 40 ml of tetrahydrofuran were dissolved 1.8 g of ethyl2-(4-hydroxyphenyl)-2-oxoacetate, 1.74 g of(3R)-1-tert-butoxycarbonyl-3-hydroxypyrrolidine and 2.92 g oftriphenylphosphine. At room temperature, 1.94 g of diethylazodicarboxylate was added to the above solution, and the resultingmixture was stirred for 18 hours. After distilling off the solvent, theresidue thus obtained was dissolved in ethyl acetate, and the solutionwas washed with water, and then dried. Thereafter, the solvent wasdistilled-off, and the resulting residue was purified by silica gelcolumn chromatography using a toluene/chloroform mixture as an eluant,thereby obtaining 2.53 g of the title compound as a viscous yellow oil.

¹ H-NMR (CDCl₃) δ:1.41 (3H, t, J=7.0Hz), 1.46 (9H, s), 2.00-2.40 (2H,m), 3.00-3.75 (4H, m), 4.43 (2H, q, J=7.0Hz), 5.00 (1H, br), 6.93 (2H,d, J=9.0Hz), 8.00 (2H, d, J=9.0Hz)

The following compounds of Reference Examples 14 to 25 were prepared inthe same manner as described in Reference Example 13.

REFERENCE EXAMPLE 14 methyl2-4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-2-oxoacetate

viscous yellow oil

REFERENCE EXAMPLE 15 ethyl2-[4-[((3R)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-2-oxoacetate

viscous yellow oil

¹ H-NMR (CDCl₃) δ: 1.40 (3H, t, J=7.0Hz), 1.46 (9H, s), 2.00-2.35 (2H,m), 3.45-3.75 (4H, m), 4.40 (2H, q, J=7.0Hz), 4.9-5.1 (1H, br), 6.95(2H, d, J=9.0Hz), 8.00 (2H, d, J=9.0Hz)

REFERENCE EXAMPLE 16 ethyl2-[4-[((2S)-1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxy]phenyl]-2-oxoacetate

viscous yellow oil

¹ H-NMR (CDCl₃) δ:1.41 (3H, t), 1.47 (9H, s), 2.0 (4H, br), 3.37 (2H,br), 4.20 (3H, br), 4.43 (2H, q), 7.0 (2H, d), 7.95 (2H, d)

REFERENCE EXAMPLE 17 ethyl2-[4-[((2S,4S)-1-tert-butoxycarbonyl-2-carbamoyl-4-pyrrolidinyl)oxy]phenyl]-2-oxoacetate

viscous yellow oil

¹ H-NMR (CDCl₃) δ:1.42 (3H, t, J=7.0Hz), 1.48 (9H, s), 2.20-2.90 (2H,br), 3.64-3.90 (2H, br), 4.30-4.60 (1H, br), 4.42 (2H, q, J=7.0Hz), 5.06(1H, br), 6.97 (2H, d, J=9.0Hz) , 8.07 (2H, d, J=9.0Hz)

REFERENCE EXAMPLE 18 ethyl 2-[4-[((2S,4S)-1-tert-butoxycarbonyl-2-dimethyl-carbamoyl-4-pyrrolidinyl)oxy]phenyl]-2-oxoacetate

viscous yellow oil

¹ H-NMR (CDCl₃) δ: 1.37-1.50 (12H, m), 1.96-2.30 (1H, m), 2.50-2.82 (1H,m), 2.90-3.15 (6H, br), 3.70 (1H, dd, J=10.8 and 5.1Hz), 3.90-4.16 (1H,m), 4.46 (2H, q, J=7.0Hz), 4.60-5.14 (2H, m), 7.00 (2H, d, J=9.4Hz),8.08 (2H, d, J=9.4Hz)

REFERENCE EXAMPLE 19 ethyl2-[4-[2-(tert-butoxycarbonylamino)-1-(tert-butoxycarbonylaminomethyl)ethoxy]phenyl]-2-oxoacetate

viscous yellow oil

¹ H-NMR (CDCl₃) δ: 1.00-1.70 (21H, br), 2.80-3.80 (4H, m), 4.20-4.60(3H, m), 7.10 (2H, d, J=8.3Hz), 7.98 (2H, d, J =8.3Hz)

REFERENCE EXAMPLE 20 ethyl2-[4-[(1-tert-butoxycarbonyl-4-piperidinyl)oxy]phenyl]-2-oxoacetate

viscous yellow oil

¹ H-NMR (CDCl₃) δ: 1.35 (3H, t, J=6Hz), 1.49 (9H, s), 1.8-2.0 (4H, m),3.2-4.0 (4H, m), 4.46 (2H, q, J=6Hz), 4.6-4.8 (1H, m), 7.01 (2H, d,J=9Hz), 8.04 (2H, d, J=9Hz)

REFERENCE EXAMPLE 21 ethyl2-[4-(2-tert-butoxycarbonylaminoethoxy)phenyl]-2-oxoacetate

viscous yellow oil

¹ H-NMR (CDCl₃) δ:1.42 (3H, t, J=7.0Hz), 1.46 (9H, s), 3.56 (2H, q,J=5.4Hz), 4.12 (2H, quintet, J=5.4Hz), 4.44 (2H, q, J=7.0Hz), 5.04 (1H,br), 6.98 (2H, d, J=9.0Hz), 8.00 (2H, d, J=9.0Hz)

REFERENCE EXAMPLE 22 ethyl2-[4-[(1-tert-butoxycarbonyl-4-piperidinyl)methoxy]phenyl]-2-oxoacetate

viscous yellow oil

¹ H-NMR (CDCl₃) δ: 1.2-1.3 (2H, m), 1.42 (3H, t, J=7.1Hz), 1.47 (9H, s),1.65-1.80 (2H, m), 3.89 (2H, d), 4.10-4.25 (2H, m), 4.43 (2H, q,J=7.1Hz), 6.95 (2H, d, J=8.8Hz), 7.99 (2H, d, J=8.8Hz)

REFERENCE EXAMPLE 23 ethyl2-[4-[((2S)-1-tert-butoxycarbonyl-5-oxo-2-pyrrolidinyl)methoxy]phenyl]-2-oxoacetate

viscous yellow oil

¹ H-NMR (CDCl₃) δ: 1.35 (3H, t), 1.41 (9H), 1.80-2.20 (2H, m), 2.47 (2H,t), 4.05 (2H, br), 4.41 (2H, q), 4.70-5.00 (1H, m), 6.98 (2H, d), 8.00(2H, d)

REFERENCE EXAMPLE 24 ethyl2-[4-[((2R,4S)-1-tert-butoxycarbonyl-2-methyl-4-pyrrolidinyl)oxy]phenyl]-2-oxoacetate

viscous yellow oil

¹ H-NMR (CDCl₃) δ:1.20-1.42 (6H, m), 1.47 (9H, s), 2.20-2.60 (1H, m),3.50-3.80 (2H, m), 3.90-4.22 (1H, m), 4.42 (2H, q), 4.90-5.10 (1H, m),6.95 (2H, d), 8.00 (2H, d)

REFERENCE EXAMPLE 25 methyl2-oxo-2-[4-[((3)-tetrahydro-3-furanyl)oxy]phenyl]acetate

Viscous yellow oil

REFERENCE EXAMPLE 26 ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-2-ethoxycarbonylacetate

a) 27.7 g of ethyl 4-methoxyphenylacetate and 34 ml of diethyl carbonatewere dissolved in 150 ml of N,N-dimethylformamide, and the solution wassubjected to reflux under heating, while gradually adding 6.5 g ofsodium hydride for 1 hour. After further refluxing under heating for 2hours, the resulting reaction solution was poured into a mixture of icewater and hydrochloric acid, followed by extraction with ethyl acetate.The resulting organic layer was washed with water, and then dried todistill off the solvent. The residue thus obtained was purified bysilica gel column chromatography using toluene as an eluant, therebyobtaining 26.7 g of ethyl 2-ethoxycarbonyl-2-(4-methoxyphenyl)acetate inthe form of light yellow oil.

¹ H-NMR (CDCl₃) δ:1.25 (6H, t, J=7.0Hz), 3.79 (3H, s), 4.20 (4H, q,J=7.0Hz), 4.55 (1H, s), 6.88 (2H, d, J=8.0Hz), 7.32 (2H, d, J=8.0Hz)

b) 5.8 g of ethyl 2-ethoxycarbonyl-2-(4-methoxyphenyl)acetate obtainedin the above step a) was dissolved in 70 ml of dichloromethane, and thesolution was cooled down to -40° C. With stirring, to this was addeddropwise 6.2 ml of boron tribromide dissolved in 5 ml ofdichloromethane. After completion of the dropwise addition, the solutionwas warmed up to room temperature, and stirred for 30 minutes. Theresulting reaction solution was poured into a mixture of ice water andhydrochloric acid, followed by extraction with chloroform. The resultingorganic layer was dried to distill off the solvent, and the residue thusobtained was purified by silica gel column chromatography usingchloroform as an eluant, thereby obtaining 4.7 g of ethyl2-ethoxycarbonyl-2-(4-hydroxyphenyl)acetate in the form of colorlessoil.

¹ H-NMR (CDCl₃) δ:1.27 (6H, t, J=7.0Hz), 4.22 (4H, q, J=7.0Hz), 4.55(1H, s), 5.66 (1H, br), 6.76 (2H, d, J=8.0Hz), 7.25 (2H, d, J=8.0Hz)

c) In 150 ml of tetrahydrofuran were dissolved 4.7 g of ethyl2-ethoxycarbonyl-2-(4-hydroxyphenyl)acetate obtained in the above stepb), 6.58 g of triphenylphosphine and 4.7 g of(3R)-1-tert-butoxycarbonyl-3-hydroxypyrrolidine. With stirring, 4.37 gof diethyl azodicarboxylate was added to the thus prepared solution, andthe stirring was continued for 18 hours. After distilling-off thesolvent, the resulting residue was purified by silica gel columnchromatography using a toluene/ethyl acetate mixture as an eluant,thereby obtaining 4.0 g of the title compound in the form of colorlessoil.

¹ H-NMR (CDCl₃) δ:1.25 (6H, t, J=7.0Hz), 1.46 (9H, s), 2.1 (2H, br),3.55 (4H, br), 4.20 (4H, q, J=7.0Hz), 4.52 (1H, s), 4.82 (1H, br), 6.82(2H, d, J=8.0Hz), 7.28 (2H, d, J =8.0Hz)

REFERENCE EXAMPLE 27 ethyl2-[4-[((2R)-1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxy]phenyl]-2-ethoxycarbonylacetate

This compound was prepared in accordance with the procedure described inReference Example 26.

viscous oil

¹ H-NMR (CDCl₃) δ:1.25 (6H, t, J=7.0Hz), 1.47 (9H, s), 2.0 (4H, br),3.40 (2H, br), 3.9 (1H), 4.20 (6H), 4.54 (1H, s), 6.82 (2H, d, J=8.0Hz),7.28 (2H, d, J=8.0Hz)

REFERENCE EXAMPLE 28 ethyl2-ethoxycarbonyl-2-[4-[(2-imidazolin-2-yl)methoxy]phenyl]acetate

a) To 150 ml of acetone were added 14.58 g of ethyl2-ethoxycarbonyl-2-(4-hydroxyphenyl)acetate, 8.8 g of bromoacetonitrileand 9.6 g of anhydrous potassium carbonate. After refluxing underheating for 5 hours, insoluble materials were removed by filtration, andthe resulting filtrate was concentrated to dryness. The residue thusobtained was purified by silica gel column chromatography using tolueneas an elution solvent, thereby obtaining 14.2 g of ethyl2-[4-(cyanomethoxy)phenyl]-2-ethoxycarbonylacetate in the form ofcolorless oil.

¹ H-NMR (CDCl₃) δ:1.26 (6H, t, J=8.0Hz), 4.22 (4H, q, J=8.0Hz), 4.58(1H, s), 4.75 (2H, s), 7.02 (2H, d, J=9.0Hz), 7.36 (2H, d, J=9.0Hz)

b) 14.2 g of ethyl 2-[4-(cyanomethoxy)phenyl]-2-ethoxycarbonylacetateobtained in the above step a) was dissolved in a mixture consisting of20 ml of ethanol and 150 ml of diethyl ether. The resulting solution wasstirred at room temperature for 18 hours with ice cooling in a blowingstream of hydrogen chloride. By distilling-off the solvent, 16.9 g ofethyl 2-ethoxycarbonyl-2-[4-(2-ethoxy-2-iminoethoxy)phenyl]acetatehydrochloride in a solid form.

c) With ice cooling and stirring, 40 ml of ethanol solution containing3.6 g of ethyl2-ethoxycarbonyl-2-[4-(2-ethoxy-2-iminoethoxy]phenyl]acetate obtained inthe above step b) was added dropwise to a 10 ml ethanol solutioncontaining 0.6 g of ethylenediamine, and the resulting mixture wasstirred at room temperature for 1.5 hours, followed by refluxing underheating for 0.5 hours. After cooling, the resulting reaction solutionwas adjusted to an acidic pH with ethanol containing 13% (w/v) ofhydrochloric acid, and then concentrated to dryness. The residue thusobtained was dissolved in water and washed with diethyl ether.Thereafter, the resulting water layer was adjusted to pH 9-10 withdilute sodium hydroxide aqueous solution, and crystals thus precipitatedwere collected by filtration. In this way, 1.83 g of the title compoundwas obtained in the form of colorless crystals.

mp: 72-110° C. (gradual wetting)

FAB MS (m/z): 335 (M⁺ +1)

¹ H-NMR (CDCl₃) δ: 1.23 (6H, t, J=8.0Hz), 3.62 (4H, s), 4.10 (4H, q,J=8.0Hz), 4.52 (1H, s), 4.68 (2H, s), 6.94 (2H, d, J=10.0Hz), 7.26 (2H,d, J=10.0Hz)

REFERENCE EXAMPLE 29 Preparation of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-benzofuranyl)propionate

a) 3.12 g of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-2-oxoacetatewas dissolved in 100 ml of tetrahydrofuran, followed by the addition of4.65 g of (5-cyano-2-benzofuranyl)methyltriphenylphosphonium chloride.To the thus prepared solution was added 400 mg of 60% sodium hydride.With stirring, to the resulting mixture was added dropwise 3 ml ofethanol, followed by stirring at room temperature for 1 hour. Theresulting reaction solution was neutralized with 10% citric acidsolution, extracted with ethyl acetate, and then dried to distill offthe solvent. Thereafter, the residue thus obtained was subjected tosilica gel column chromatography using a toluene/ethyl acetate mixtureas an elution solvent, thereby obtaining 3.1 g of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-benzofuranyl)acrylatein the form of viscous oil as a mixture of E and Z forms. A portion ofthe thus obtained compound was separated into E and Z forms.

E form (less polar):

¹ H-NMR (CDCl₃) δ: 1.32 (3H, t, J=7.6Hz), 1.49 (9H, s), 1.70-2.40 (2H,m), 3.30-3.80 (4H, m), 4.30 (2H, q, J=7.6Hz), 4.92 (1H, br), 6.62 (1H,s), 6.94 (2H, d, J=9.0Hz), 7.24 (2H, d, J=9.0Hz), 7.38 (1H, d, J=8.6Hz),7.56 (1H, d, J=8.6Hz), 7.74 (1H, s), 7.77 (1H, s)

Z form:

¹ H-NMR (CDCl₃) δ: 1.10-1.60 (12H, m), 2.00-2.30 (2H, m), 3.30-3.80 (4H,m), 4.50 (2H, q, J=7.2Hz), 4.92 (1H, br), 6.76 (1H, s), 6.81 (1H, s),6.88 (2H, d, J=8.75Hz), 7.88 (2H, d, J=8.75Hz), 7.31-7.60 (2H), 7.85(1H, s)

b) 3.1 g of ethyl2-[4-[((3S)-1-tert-butoxy-carbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-benzofuranyl)acrylateobtained in the above step a) was dissolved in a mixed solvent system of100 ml of tetrahydrofuran and 100 ml of ethanol. To this was added 700mg of palladium oxide.1H₂ O.barium sulfate which had been prepared inaccordance with the procedure disclosed in Angewandte Chemie, vol.67,p.785, 1955. After catalytic hydrogenation under normal pressure for 6hours, the catalyst was removed by filtration, and the resultingfiltrate was concentrated. Thereafter, the thus obtained residue wassubjected to silica gel column chromatography using a toluene/ethylacetate mixture as an elution solvent, thereby obtaining 1.9 g of thetitle compound as a viscous oil.

¹ -NMR (CDCl₃) δ: 1.00-1.40 (3H, m), 1.46 (9H, s), 2.00-2.30 (2H, m),3.16 (1H, dd, J=14.4 and 7.2Hz), 3.40-3.80 (5H, m), 3.90-4.30 (3H, m),4.94 (1H, br), 6.40 (1H, s), 6.80 (2H, d, J=8.7Hz), 7.25 (2H, d,J=8.7Hz), 7.46 (2H, s), 7.76 (1H, s)

REFERENCE EXAMPLE 30 Preparation of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-cyano-2-naphthyl)propionate

a) 8.40 g of (7-cyano-2-naphthyl)methyltriphenylphosphoniumbromide and5.0 g of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-2-oxoacetatewere suspended in a mixture of 100 ml of tetrahydrofuran and 100 ml ofethanol. With stirring, to the resulting suspension was added 2.51 g of1,8-diazabicyclo[5.4.0]-7-undecene, followed by stirring for 3 hours atroom temperature. After distilling off the solvent, the residue thusobtained was subjected to silica gel column chromatography using an-hexane/ethyl acetate mixture as an elution solvent, thereby obtaining6.06 g of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-cyano-2-naphthyl)acrylateas a mixture of E and Z forms. A portion of the thus obtained compoundwas separated into E and Z forms.

E form:

mp: 104°-106° C. (crystallization in ethanol)

¹ H-NMR (CDCl₃) δ: 1.35 (3H, t, J=7.3Hz), 1.48 (9H, s), 2.05-2.30 (2H,m), 3.45-3.70 (4H, m), 4.31 (2H, q, J=7.3Hz), 4.92 (1H, br), 6.86 (2H,d, J=8.8 Hz), 7.16 (2H, d, J=8.8Hz), 7.20 (1H, dd, J=8.8 and 1.5 Hz),7.56 (1H, dd, J=8.3 and 1.5Hz), 7.62 (1H, d, J=8.8 Hz), 7.73 (1H, s),7.80 (1H, d, J=8.3Hz), 7.93 (1H, s), 8.07 (1H, s)

Z form:

¹ H-NMR (CDCl₃) δ:1.19 (3H, t, J=7.3Hz), 1.48 (9H, s), 2.05-2.30 (2H,m), 3.45-3.70 (4H, m), 4.29 (2H, q, J=7.3Hz), 4.93 (1H, br), 6.90 (2H,d, J=8.8Hz), 7.09 (1H, s), 7.44 (2H, d), 7.60 (1H, dd, J=8.3 and 1.5Hz),7.63 (1H, dd, J=8.8 and 1.5Hz), 7.85 (1H, d, J=8.8Hz), 7.88 (1H, s),7.90 (1H, d, J=8.3Hz), 8.18 (1H, s)

b) 6.06 g of ethyl2-[4-[((3S)-1-tert-butoxy-carbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-cyano-2-naphthyl)acrylateobtained as a mixture of E and Z forms in the above step a) wasdissolved in a mixed solvent system of 80 ml of tetrahydrofuran and 80ml of ethanol. To this was added 2.0 g of palladium oxide.1H₂ O.bariumsulfate. After catalytic hydrogenation under normal pressure for 3.5hours, the catalyst was removed by filtration, and the solvent wasdistilled off. Thereafter, the thus obtained residue was subjected tosilica gel column chromatography using a n-hexane/ethyl acetate mixtureas an elution solvent, thereby obtaining 6.24 g of the title compound ina partially solidified form.

¹ H-NMR (CDCl₃) δ:1.11 (3H, t, J=7.3Hz), 1.47 (9H, s), 2.00-2.33 (2H,m), 3.18 (1H, dd, J=14.2 and 6.8Hz), 3.40-3.65 (5H, m), 3.88 (1H, t,J=7.5Hz), 4.06 (2H, q, J=7.3Hz), 4.85 (1H, br), 6.80 12H, d, J=8.8Hz),7.24 (2H, d), 7.42 (1H, dd, J=8.8 and 1.5Hz), 7.54 (1H, dd, J=8.3 and1.5Hz), 7.62 (1H, s), 7.77 (1H, d, J=8.8Hz), 7.85 (1H, d, J=8.3Hz), 8.13(1H, s)

The following compounds of Reference Examples 31 to 39 were prepared inaccordance with the procedure described in Reference Example 30.

REFERENCE EXAMPLE 31 ethyl2-[4-[((3R)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-cyano-2-naphthyl)propionate

¹ H-NMR (CDCl₃) δ:1.11 (3H, t, J=7.3Hz), 1.47 (9H, s), 2.00-2.35 (2H,m), 3.18 (1H, dd, J=14.2 and 6.8Hz), 3.40-3.70 (5H, m), 3.88 (1H, br),4.06 (2H, q, J=7.3Hz), 4.85 (1H, br), 6.80 (2H, d, J=8.8Hz), 7.24 (2H),7.42 (1H, dd, J=8.8 and 1.5Hz), 7.54 (1H, dd, J=8.3 and 1.5Hz), 7.62(1H, s), 7.77 (1H, d, J=8.8Hz), 7.84 (1H, d, J=8.3Hz), 8.11 (1H, s)

REFERENCE EXAMPLE 32 ethyl 2-[4-[(1-tert-butoxycarbonyl-4-piperidinyl)oxy]phenyl]3-(7-cyano-2-naphthyl)propionate

¹ H-NMR (CDCl₃) δ:1.11 (3H, t), 1.49 (9H, s), 1.70-2.00 (4H, m),3.00-4.10 (9H, m), 4.45 (1H, br), 6.80-8.10 (10H, m)

FAB MS (m/z): 418 (M⁺ +1)

REFERENCE EXAMPLE 33 ethyl2-[4[((2S,4S)-1-tert-butoxycarbonyl-2-carbamoyl-4-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-benzofuranyl)propionate

viscous oil

¹ H-NMR (CDCl₃) δ: 1.16 (3H, t, J=7.0Hz), 1.47 (9H, s), 2.10-2.80 (2H,br), 3.16 (1H, dd, J=14.4 and 7.2Hz), 3.40-4.50 (6H, m), 5.08 (1H, br),5.80 (1H, br), 6.39 (1H, s), 6.76 (2H, d, J=8.35Hz), 7.26 (2H, d,J=8.35Hz), 7.50 (2H, s), 7.80 (1H)

REFERENCE EXAMPLE 34 ethyl2-[4-[((2S,4S)-1-tert-butoxycarbonyl-2-dimethylcarbamoyl-4-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-benzofuranyl)propionate

viscous oil

¹ H-NMR (CDCl₃) δ:1.23 (3H, t), 1.44 (9H, s), 1.90-2.30 (1H, br),2.40-2.80 (1H, br), 2.98 (1H, s), 3.10 -4.23 (7H, m), 4.40-5.00 (2H,br), 6.38 (1H, s), 6.90 (2H, d, J=8.3Hz), 7.20 (2H, d, J=8.35Hz), 7.45(2H, s), 7.76 (1H, s)

REFERENCE EXAMPLE 35 ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]3-(5-cyano-3-methyl-2-benzofuranyl)propionate

viscous oil

¹ H-NMR (CDCl₃) δ: 1.16 (3H, t), 1.47 (9H, s), 2.02 (3H, s), 2.1 (2H,br), 3.1 (1H, br), 3.6 (5H, br), 4.1 (3H, m), 4.85 (1H, br), 6.83 (2H,d), 7.15 (2H, d), 7.46 (2H), 7.7 (1H, s)

REFERENCE EXAMPLE 36 ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-7-methoxy-2-benzofuranyl)propionate

viscous yellow oil

¹ H-NMR (CDCl₃) δ:1.17 (3H, t, J=7Hz), 1.46 (9H, s), 2.00-2.30 (2H, m),3.16 (1H, dd, J=14.5 and 7.4Hz), 3.40-3.76 (5H, m), 3.80-4.30 (3H, m),4.02 (3H, s), 4.70-5.00 (1H, br), 6.37 (1H, s), 6.80 (2H, d, J=8.75Hz),6.95 (1H, d, J=1.3Hz), 7.23 (2H, d, J=8.75Hz), 7.41 (1H, d, J=1.3Hz)

REFERENCE EXAMPLE 37 ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-3-benzofuranyl)propionate

viscous oil

¹ H-NMR (CDCl₃) δ:1.14 (3H, t), 1.45 (9H, s), 2.12 (2H, br), 2.90-4.00(7H, m), 4.08 (2H, q), 4.84 (1H, br), 6.85 (2H, d), 7.2 (2H, d), 7.41(1H, s), 7.50 (2H), 7.72 (1H)

REFERENCE EXAMPLE 38 ethyl 2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyano-2-naphthyl)propionate

¹ H-NMR (CDCl₃) δ: 1.25 (3H, t, J=7.0Hz), 1.46 (9H, s), 2.00-2.20 (2H,m), 3.00-4.00 (7H, m), 4.08 (2H, q), 4.85 (1H, br), 6.80-8.20 (10H, m)

REFERENCE EXAMPLE 39 ethyl2-[4-[(1-tert-butoxycarbonyl-4-piperidinyl)methoxy]phenyl]-3-(7-cyano-2-naphthyl)propionate

¹ H-NMR (DMSO-d₆) δ:1.01 (3H, t, J=7.1Hz), 1.1-1.2 (2H, m), 1.39 (9H,s), 1.68-1.76 (2H, m), 2.65-2.75 (2H, m), 3.78 (2H, d), 3.9-4.1 (5H, m),4.55-4.65 (1H, m), 6.85 (2H, d, J=8.3Hz), 7.25 (2H, d, J=8.3Hz),7.55-7.65 (1H, m), 7.68-7.73 (1H, m), 7.82 (1H, s), 7.90-7.95 (1H, m),8.03 (1H, d, J=8.8Hz), 8.44 (1H, s)

REFERENCE EXAMPLE 40 Preparation of ethyl(+)-2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-cyano-2-naphthyl)propionateand ethyl(-)-2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-cyano-2-naphthyl)propionate

2.0 g of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-cyano-2-naphthyl)propionatewas dissolved in 10 ml of ethanol under warming. After cooling to roomtemperature, crystals thus precipitated were collected by filtration,and then recrystallized from ethanol twice to obtain 640 mg of ethyl(+)-2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-cyano-2-naphthyl)propionate.

mp: 132°-133.5° C.

[α]_(D) ²⁴ =+117.4° (c=1.008, CHCl₃)

¹ H-NMR (CDCl₃) δ:1.11 (3H, t, J=7.3Hz), 1.47 (9H, s), 2.00 -2.30 (2H,m), 3.18 (1H, dd, J=14.2 and 6.8Hz), 3.40-3.70 (5H, m), 3.87 (1H, t,J=7.6Hz), 4.00-4.10 (2H, m), 4.85 (1H, br), 6.80 (2H, d, J=8.8Hz),7.20-7.30 (2H, m), 7.42 (1H, d, J=8.3Hz), 7.55 (1H, .d, J=8.3Hz), 7.63(1H, s), 7.77 (1H, d, J=8.3Hz), 7.85 (1H, d, J=8.3Hz), 8.12 (1H, s)

HPLC: Column; an amylose-based column for use in the separation ofoptical isomers (CHIRALPAK AD, 4.66 φ×250 mm, Daicel ChemicalIndustries, Ltd.)

Solvent; iso-propanol:n-hexane=15:85 (v/v)

Flow rate; 1 ml/min

Column temperature; 25° C.

Retention time; 31.37 minutes

The filtrate was concentrated to dryness and crystallized from an-hexane/ethanol mixture. The crystals thus collected wererecrystallized from the same mixed solvent system to obtain 80 mg ofethyl(-)-2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-cyano-2-naphthyl)propionate.

mp 82.5°-85.0° C.

[α]_(D) ²⁴ +-85.0° (c =0.53, CHCl₃)

¹ H-NMR (CDCl₃) δ:1.11 (3H, t, J=7.3Hz), 1.47 (9H, s), 2.00-2.30 (2H,m), 3.18 (1H, dd, J=14.2 and 6.8Hz), 3.40-3.66 (5H, m), 3.87 (1H, t,J=7.6Hz), 4.00-4.10 (2H, m), 4.85 (1H, br), 6.80 (2H, d, J=8.8Hz),7.20-7.30 (2H, m), 7.42 (1H, d, J=8.3Hz), 7.56 (1H, d, J=8.3Hz), 7.62(1H, s), 7.77 (1H, d, J=8.3Hz), 7.85 (1H, d, J=8.3Hz), 8.12 (1H, s)

HPLC: Column; an amylose-based column for use in the separation ofoptical isomers (CHIRALPAK AD, 4.6φ×250 mm, Daicel Chemical Industries,Ltd.)

Solvent; iso-propanol:n-hexane=15:85 (v/v)

Flow rate; 1 ml/min

Column temperature; 25° C.

Retention time; 23.22 minutes

REFERENCE EXAMPLE 41 Preparation of ethyl3-(5-cyano-2-benzofuranyl)-2-[4-[(3S)-1-methyl-3-pyrrolidinyl)oxy]phenyl]propionate

1.8 g of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-benzofuranyl)propionatewas dissolved in 28 ml of formic acid, and the solution was stirred at70° C. for 1 hour. The resulting reaction solution was concentrated todryness, and the residue thus obtained was dissolved in 8 ml of formicacid. 0.29 ml of 37% formaldehyde was added thereto, and then refluxedunder heating for 4 hours. After cooling, the reaction solution wasmixed with chloroform, and then adjusted to pH 10-11 with aqueousammonia, and the resulting organic layer was collected and dried.Thereafter, the solvent was distilled off, and the resulting residue waspurified by silica gel column chromatography using a chloroform/methanolmixture as an eluant. In this way, 1.07 g of the title compound wasobtained in an oily form.

¹ H-NMR (CDCl₃) δ:1.16 (3H, t, J=7.2Hz), 1.60-2.30 (2H, m), 2.38 (3H,s), 2.00-4.00 (7H, m), 4.11 (2H, q, J=7.2Hz), 4.60-4.90 (1H, br), 6.39(1H, s), 6.78 (2H, d, J=8.8Hz), 7.21 (2H, d, J=8.8 Hz), 7.47 (2H, s),7.77 (1H, s)

REFERENCE EXAMPLE 42 Preparation of ethyl2-[4-[((3S)-1-acetyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-benzofuranyl)propionate

2.3 g of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-benzofuranyl)propionatewas dissolved in 3 ml of anisole, 25 ml of trifluoroacetic acid wasadded to the above solution with ice cooling, and the resulting mixturewas stirred at room temperature for 1 hour. After distilling offtrifluoroacetic acid under reduced pressure, the residue thus obtainedwas adjusted to pH 10-11 with saturated sodium bicarbonate aqueoussolution, extracted with chloroform, and then dried. At roomtemperature, the resulting organic layer was mixed with 2 ml oftriethylamine and then with 555 mg of acetyl chloride, followed bystirring at the same temperature for 0.5 hours. After distilling off thesolvent, the resulting residue was subjected to silica gel columnchromatography using a chloroform/ethanol mixture as an eluant to obtain1.8 g of the title compound.

¹ H-NMR (CDCl₃) δ:1.17 (3H, t, J=7.0Hz), 2.04 (1.5H) 2.08 (1.5H), 3.14(1H, dd, J=15.1 and 3.6Hz), 3.40-4.30 (8H, m), 4.70-5.04 (1H, br), 6.40(1H, s), 6.60-6.92 (2H, m), 7.10 (2H, m), 7.47 (2H, s), 7.77 (1H, s)

REFERENCE EXAMPLE 43 Preparation of ethyl3-(5-cyano-2-benzofuranyl)-2-[4-[((3S)-1-dimethylcarbamoyl-3-pyrrolidinyl)oxy]phenyl]propionate

2.3 g of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-benzofuranyl)propionatewas dissolved in 3 ml of anisole. 25 ml of trifluoroacetic acid wasadded to the above solution with ice cooling, and the resulting mixturewas stirred at room temperature for 1 hour. After distilling offtrifluoroacetic acid under reduced pressure, the residue thus obtainedwas adjusted to pH 10-11 with saturated sodium bicarbonate aqueoussolution, extracted with chloroform, and then dried. At roomtemperature, the resulting organic layer was mixed with 2 ml oftriethylamine and then with 760 mg of N,N-dimethylcarbamoyl chloride,followed by stirring at the same temperature for 1 hour. Afterdistilling off the solvent, the resulting residue was subjected tosilica gel column chromatography using a chloroform/ethanol mixture asan eluant to obtain 1.7 g of the title compound.

¹ H-NMR (CDCl₃) δ: 1.17 (3H, t, J=7.0Hz), 1.9-2.20 (2H, m) 2.86 (6H, s),3.14 (1H, dd, J=16.0 and 7.2Hz), 3.30-4.50 (8H, m), 4.72-4.96 (1H,br),6.41(1H, s), 6.83 (2H, d, J =8.7Hz), 7.25 (2H, d, J=8.7Hz), 7.49(2H, s), 7.78 (1H, s)

REFERENCE EXAMPLE 44 Preparation of ethyl2-(4-acetoxyphenyl)-2-oxoacetate

7.25 g of ethyl 2-(4-hydroxyphenyl)-2-oxoacetate was dissolved in 15 mlof pyridine, followed by adding 4 ml of acetic anhydride and thenstirring at room temperature for 1 hour. The resulting reaction solutionwas poured into water, and extracted with diethyl ether. The resultingorganic layer was washed with water, and then concentrated to dryness.Thereafter, the residue was dissolved in benzene, and then concentratedto obtain 8.3 g of the title compound in an oily form.

¹ H-NMR (CDCl₃) δ: 1.41 (3H, t), 2.32 (3H, s), 4.43 (2H, q), 7.29 (2H,d), 8.01 (2H, d)

REFERENCE EXAMPLE 45 Preparation of ethyl3-(5-cyano-2-benzofuranyl)-2-(4-hyroxyphenyl)propionate

a) 15.93 g of (5-cyano-2-benzofuranyl)methyltriphenylphosphoniumchloride and 8.29 g of ethyl 2-(4-acetoxyphenyl)-2-oxoacetate weredissolved in a mixture solution of 80 ml of tetrahydrofuran and 80 ml ofethanol. At room temperature, 5.34 g of1,8-diazabicyclo[5.4.0]-7-undecene was added to the above solution, andthe mixture was stirred for 18 hours at the same temperature. Theresulting reaction solution was concentrated to dryness, and the residuewas purified by silica gel column chromatography using a toluene/ethylacetate mixture as an eluant to obtain 11.28 g of ethyl2-(4-acetoxyphenyl)-3-(5-cyano-2-benzofuranyl)acrylate in the form oflight yellow crystals as a mixture of E and Z forms.

¹ H-NMR (CDCl₃) δ: 1.32 (3H, t), 2.36 (3H, s), 4.30 (2H, q), 6.30 (1H,s), 7.2-7.8 (8H, m)

b) 3.8 g of ethyl 2-(4-acetoxyphenyl)-3-(5-cyano-2-benzofuranyl)acrylateobtained in the above step a) was dissolved in anethanol/tetrahydrofuran mixture solvent. The resulting solution wasmixed with 750 mg of palladium oxide.1H₂ O.barium sulfate and subjectedto catalytic hydrogenation under normal pressure. After removing thecatalyst by filtration, the resulting filtrate was concentrated todryness to obtain 3.8 g of ethyl2-(4-acetoxyphenyl)-3-(5-cyano-2-benzofuranyl)propionate.

¹ H-NMR (CDCl₃) δ:1.16 (3H, t, J=7.2Hz), 2.25 (3H, s), 3.20 (1H, dd,J=16.2 and 7.0Hz), 3.40-4.30 (4H, m), 6.50 (1H, s), 7.10 (2H, d,J=8.3Hz), 7.40 (2H, d, J=8.3Hz), 7.56 (2H, s), 7.86 (1H, s)

c) 8.1 g of ethyl2-(4-acetoxyphenyl)-3-(5-cyano-2-benzofuranyl)propionate obtained in theabove step b) was dissolved in 100 ml of ethanol solution containing 15%of ammonia, and the resulting solution was allowed to stand still for 18hours at room temperature. Thereafter, the resulting reaction solutionwas concentrated to dryness, and the residue was purified by silica gelcolumn chromatography using chloroform as an eluant. In this way, 5.62 gof the title compound was obtained in the form of colorless crystals.

mp: 140°-142° C.

¹ H-NMR (CDCl₃) δ:1.15 (3H, t), 3.0-4.0 (3H, m), 4.1 (2H, q), 4.98 (1H,s), 6.39 (1H, s), 6.76 (2H, d), 7.15 (2H, d), 7.45 (2H), 7.75 (1H)

REFERENCE EXAMPLE 46 Preparation of ethyl3-(5-cyano-2-benzofuranyl)-3-(4-hydroxyphenyl)propionate

a) To 150 ml of acetone were suspended 20 g of 5-bromosalicylaldehyde,22.9 g of 2-bromo-4-methoxyacetophenone and 27.6 g of anhydrouspotassium carbonate. After stirring at room temperature for 4 hours, theresulting reaction solution was concentrated to dryness, and then mixedwith water to collect precipitated crystals by filtration. After washingwith water and subsequent recrystallization from ethanol, 14.02 g of5-bromo-2-(4-methoxybenzoyl)benzofuran was obtained in the form ofcolorless prism crystals.

mp: 143°-146° C.

IR (KBr): 1644, 1605, 1257 cm⁻¹

¹ H-NMR (DMSO-d₆) δ:3.35 (3H, s), 7.15 (2H, d, J=9Hz), 7.72 (3H, m),8.0-8.2 (3H)

b) 15.0 g of 5-bromo-2-(4-methoxybenzoyl)benzofuran obtained in theabove step a) and 6.09 g of cuprous cyanide were suspended in 75 ml ofN-methyl-2-pyrrolidone, and the suspension was stirred at 200° to 220°C. for 5 hours in a stream of nitrogen. After cooling, the reactionsolution was diluted with chloroform to remove insoluble materials byfiltration, and the resulting filtrate was washed with dilutehydrochloric acid. After drying the organic layer and subsequentconcentration under a reduced pressure, 6.60 g of2-(4-methoxybenzoyl)-5-benzofurancarbonitrile was obtained as a brownpowder.

IR (KBr): 2224, 1644 cm⁻¹

¹ H-NMR (DMSO-d₆) δ:3.30 (3H, s), 7.15 (2H, d, J=9Hz), 7.83 (1H, s),8.00 (2H, d), 8.07 (2H, d, J=9Hz), 8.42 (1H, s)

c) 1.85 g of ethyl diethylphosphonoacetate was dissolved in 20 ml oftetrahydrofuran. With stirring at room temperature, 320 mg of 60% sodiumhydride was added to the above solution, and the stirring was continueduntil the reaction solution became clear. After 10 minutes, 1.75 g of2-(4-methoxybenzoyl)-5-benzofurancarbonitrile obtained in the above stepb) was added to the above reaction solution, and the mixture wasrefluxed under heating for 30 minutes. After cooling, the resultingreaction mixture was concentrated to dryness. The residue thus obtainedwas mixed with dilute hydrochloric acid, extracted with dichloromethane,washed with saturated sodium chloride aqueous solution, and thenconcentrated to dryness. The resulting residue was purified by silicagel column chromatography using a n-hexane/dichloromethane mixture as aneluant, thereby obtaining 1.78 g of ethyl3-(5-cyano-2-benzofuranyl)-3-(4-methoxyphenyl)acrylate as a mixture of Eand Z forms.

¹ H-NMR (CDCl₃) δ:1.20 (3H, t, J=7Hz), 3.84 (3H, s), 4.18 (2H, q,J=7Hz), 6.32 (1H, s), 6.8-7.4 (5H, m), 7.56 (2H, s), 7.93 (1H, br)

d) 1.78 g of the E/Z mixture of ethyl3-(5-cyano-2-benzofuranyl)-3-(4-methoxyphenyl)acrylate obtained in theabove step c) was dissolved in a solvent mixture of 6 ml oftetrahydrofuran and 20 ml of ethanol. 200 mg of 5% palladium carboncatalyst was added to the above solution, and the mixture was subjectedto catalytic hydrogenation under normal pressure for 1.5 hours. Afterremoving the catalyst by filtration, the resulting filtrate wasconcentrated to dryness to obtain 1.79 g of ethyl3-(5-cyano-2-benzofuranyl)-3-(4-methoxyphenyl)propionate.

¹ H-NMR (CDCl₃) δ:1.29 (3H, t, J=7Hz), 2.9-3.1 (2H, m), 3.78 (3H, s),4.09 (2H, q, J=7Hz), 4.5-4.7 (1H, m), 6.47 (1H, s), 6.88 (2H, d, J=9Hz),7.24 (2H, d, J=9Hz), 7.47 (2H, s), 7.80 (1H, s)

e) 1.79 g of ethyl3-(5-cyano-2-benzofuranyl)-3-(4-methoxyphenyl)propionate obtained in theabove step d) was dissolved in 20 ml of anhydrous dichloromethane, andthe solution was cooled down to -50° C. To this solution was addeddropwise a 10 ml dichloromethane solution containing 1.36 ml borontribromide. The resulting mixture was gradually warmed up, and stirredat room temperature for 3 hours. Thereafter, the reaction solution wasdiluted with dichloromethane, and the resulting organic layer was washedwith dilute hydrochloric acid and then with saturated sodium chlorideaqueous solution, followed by concentration to dryness. In this way,1.34 g of the title compound was obtained in an oily form.

¹ H-NMR (CDCl₃) δ:1.15 (3H, t, J=7Hz), 2.9-3.3 (2H, m), 4.09 (2H, q,J=7Hz), 4.5-4.7 (1H, m), 6.15 (1H, br), 6.46 (1H, s), 6.80 (2H, d,J=9Hz), 7.15 (2H, d, J=9Hz), 7.42 (2H, s), 7.76 (1H, s)

REFERENCE EXAMPLE 47 Preparation of ethyl2-(2-(5-cyano-2-benzofuranyl)ethyl)-5-hydroxybenzoate

a) 4.87 g of 2-formyl-5-methoxybenzoic acid was dissolved in 30 ml ofchloroform. With stirring at room temperature, a benzene/n-hexane (1:1)mixture solution of diphenyldiazomethane prepared in accordance with theprocedure disclosed in Journal of the Chemical Society (Parkin I, pp.2030-2033, 1975) was added to the above solution until the reactionsolution showed a purplish red color. The resulting reaction solutionwas subjected to purification by silica gel column chromatography usinga toluene/ethyl acetate mixture as an eluant to obtain 8.2 g ofdiphenylmethyl 2-formyl-5-methoxybenzoate as a viscous oil.

¹ H-NMR (CDCl₃) δ:3.87 (3H, s), 7.13 (1H, dd, J=11.5 and 2.9Hz), 7.20(1H, s), 7.24 (11H, m), 7.97 (1H, d, J=11.5Hz), 10.45 (1H, s)

b) 6.0 g of diphenylethyl 2-formyl-5-methoxybenzoate obtained in theabove step a) and 8.1 g of(5-cyano-2-benzofuranyl)methyltriphenylphosphonium chloride weredissolved in a mixture solvent of 70 ml of tetrahydrofuran and 70 ml ofmethanol. With stirring at room temperature, 2.91 g of1,8-diazabicyclo[5.4.0]-7-undecene was added to the above solution, andthe mixture was stirred for 2 hours at thee same temperature. Afterdistilling off the solvent, the resulting residue was subjected topurification by silica gel column chromatography using atoluene/chloroform mixture as an eluant to obtain 8.2 g ofdiphenylmethyl 2-[2-(5-cyano-2-benzofuranyl)vinyl]-5-methoxybenzoate asa mixture of E and Z forms.

¹ H-NMR (CDCl₃) δ:3.84 (1H, s), 3.88 (3H, s), 6.20-8.28 (19H, m)

c) 8.2 g of the E/Z mixture of diphenylmethyl2-[2-(5-cyano-2-benzofuranyl)vinyl]-5-methoxybenzoate obtained in theabove step b) was dissolved in a solvent mixture of 60 ml oftetrahydrofuran and 60 ml of ethanol. 2.0 g of palladium oxide.1H₂O.barium sulfate was added to the above solution, and the mixture wassubjected to catalytic hydrogenation under normal pressure. Afterremoving the catalyst by filtration, the resulting filtrate wasconcentrated to collect precipitated crystals by filtration, therebyobtaining 4.45 g of 2-[2-(5-cyano-2-benzofuranyl)ethyl]-5-methoxybenzoicacid.

mp: 179°-182° C.

¹ H-NMR (CDCl₃) δ:2.90-3.42 (4H, m), 3.75 (3H, s), 6.67 (1H, s), 7.01(1H, dd, J=8.7 and 2.2Hz), 7.24 (1H, d, J=8.7Hz), 7.34 (1H, d, J=2.2Hz),7.69 (2H, s), 8.06 (1H, s), 12.98 (1H, br)

FD MS (m/z): 321 (M⁺), 311, 283

d) 4.45 g of 2-[2-(5-cyano-2-benzofuranyl)ethyl]-5-methoxybenzoic acidobtained in the above step c) was dissolved in 200 ml of ethanol, andthe solution was mixed with 4 ml of concentrated sulfuric acid, andrefluxed under heating for 16 hours. After cooling, the resultingreaction solution was neutralized with saturated sodium bicarbonateaqueous solution, and ethanol was removed by distillation. The residuethus obtained was extracted with ethyl acetate, and dried to distill offthe solvent. The resulting residue was purified by silica gel columnchromatography, and then recrystallized from n-hexane to obtain 4.11 gof ethyl 2-[2-(5-cyano-2-benzofuranyl)ethyl]-5-methoxybenzoate in theform of colorless needle crystals.

mp: 92°-93° C.

¹ H-NMR (CDCl₃) δ: 1.38 (3H, t, J=7.0Hz), 2.90-3.48 (4H, m), 3.82 (3H,s), 4.34 (2H, q, J=7.0Hz), 6.41 (1H, s), 6.96 (1H, dd, J=8.7 and 2.6Hz),7.10 (1H, d, J=8.7Hz), 7.46 (1H, d, J=2.6Hz), 7.48 (2H, s), 7.79 (1H, s)

e) 4.11 g of ethyl 2-[2-(5-cyano-2-benzofuranyl)ethyl]-5-methoxybenzoateobtained in the above step d) was dissolved in 40 ml of dichloromethane,and the solution was cooled down to -78° C. 8.85 g of boron tribromidewas added dropwise to the above solution at the same temperature, andthe mixture was gradually warmed up to -5° C. to 0° C., and stirred for1 hour. The resulting reaction solution was poured into ice water, andextracted with ethyl acetate. The resulting organic layer was washedwith 4N hydrochloric acid and then with water, followed by drying andremoval of the solvent. Thereafter, the resulting residue was purifiedby silica gel column chromatography to obtain 2.80 g of the titlecompound in the form of prism crystals.

mp: 133°-135° C.

¹ H-NMR (CDCl₃) δ:1.40 (3H, t, J=7.0Hz), 2.96-3.50 (4H, m), 4.36 (2H, q,J=7.0Hz), 6.45 (1H, s), 6.93 (1H, dd, J =8.7 and 2.9Hz), 7.13 (1H, d,J=8.7Hz), 7.50 (1H, d, J =2.9Hz), 7.56 (2H, s), 7.84 (1H, s)

REFERENCE EXAMPLE 48 Preparation of ethyl2-[2-[2-(5-cyano-2-benzofuranyl)ethyl]-5-hydroxypbenyl]acetate

a) 2.0 g of 2-[2-(5-cyano-2-benzofuranyl)ethyl]-5-methoxybenzoic acidwas suspended in 10 ml of benzene, and the suspension was mixed with 1ml of thionyl chloride. By refluxing under heating the resulting mixturefor 1 hour and subsequently concentrating to dryness, crude acidchloride was obtained.

A mixture solution consisting of 10 ml of n-hexane solution containing10% (w/v) trimethylsilyldiazomethane, 1.3 ml of triethylamine, 10 ml ofacetonitrile and 10 ml of tetrahydrofuran was cooled to -5° C. Withstirring, to this was added dropwise a 5 ml acetonitrile solution of thecrude acid chloride prepared above. The resulting reaction solution wasstirred at 0° C. for 48 hours, and the solvent was subsequentlydistilled off under a reduced pressure at a low temperature. The residuethus obtained was dissolved in a mixture solvent of 4 ml of collidineand 4 ml of benzyl alcohol, and the resulting solution was stirred at180° C. for 7 minutes in a stream of nitrogen. The resulting reactionsolution was dissolved in benzene, washed with 10% citric acid, and thendried. After distilling off the solvent, the resulting residue wassubjected to silica gel column chromatography using a toluene/ethylacetate mixture as an eluant to obtain 830 mg of benzyl2-[2-[2-(5-cyano-2-benzofuranyl)ethyl]-5-methoxyphenyl]acetate.

mp: 127°-128° C.

¹ H-NMR (CDCl₃) δ:3.00 (4H), 3.68 (2H, s), 3.76 (3H, s), 5.13 (2H, s),6.32 (1H, s), 6.76 (1H, dd, J=7.9 and 1.3Hz), 6.80 (1H, s), 7.08 (1H, d,J=7.9Hz), 7.30 (5H, s), 7.48 (1H, d, J=1.3Hz), 7.77 (1H, s)

b) 855 mg of benzyl2-[2-[2-(5-cyano-2-benzofuranyl)ethyl]-5-methoxyphenyl]acetate obtainedin the above step a) was dissolved in 20 ml of dichloromethane, and thesolution was cooled down to -50° C. To this solution was added dropwisea5 ml dichloromethane solution containing 1.75 g of boron tribromide,followed by gradually increasing temperature to 15° C. and stirring atthe increased temperature for 20 minutes. The resulting reactionsolution was extracted with ethyl acetate, washed with dilutehydrochloric acid, and then dried. After concentration to dryness, theresulting residue was dissolved in 30 ml of ethanol, mixed with 2 ml ofthionyl chloride, and then refluxed under heating for 1 hour. Aftercooling, the reaction solution was diluted with ethyl acetate, and theresulting organic layer was washed with water, and dried to distill ofthe solvent. The thus obtained residue was purified by subjecting it tosilica gel column chromatography using chloroform as an eluant. In thisway, 680 mg of the title compound was obtained in the form of powder.

mp: 84°-86° C.

¹ H-NMR (CDCl₃) δ:1.25 (3H, t, J=7.0Hz), 3.02 (4H), 3.59 (2H, s), 4.57(2H, q, J=7.0Hz), 6.19 (1H, s), 6.41 (1H, s), 6.55-6.84 (2H, m), 6.99(1H, d, J=7.9Hz), 7.48 (2H, s), 7.77 (1H, s)

REFERENCE EXAMPLE 49 Preparation of ethyl5-cyano-2-[2-(4-hydroxyphenyl)ethyl]-3-benzofurancarboxylate

a) 91.5 g of (5-bromo-2-benzofuranyl)methyltriphenylphosphonium chlorideand 25 g of p-anisaldehyde were dissolved in a mixture solvent of 180 mlof tetrahydrofuran and 180 ml of ethanol. With stirring at roomtemperature, 27.58 g of 1,8-diazabicyclo[5.4.0]-7-undecene was added tothe above solution, and the mixture was stirred for 18 hours.Thereafter, the reaction solution was concentrated to collectprecipitated crystals by filtration, thereby obtaining 32.8 g of5-bromo-2-[2-(4-methoxyphenyl)vinyl]benzofuran as one of thestereoisomers.

mp: 190°-194° C.

¹ H-NMR (CDCl₃) δ:3.83 (3H, s), 6.54 (1H, s), 6.9 (3H), 7.25 (1H, d,J=17Hz), 7.31 (2H), 7.45 (2H, d), 7.62 (1H)

The filtrate obtained above was concentrated to dryness, and theresulting residue was purified by subjecting it to silica gel columnchromatography using toluene as an eluant, thereby obtaining 22 g of5-bromo-2-[2-(4-methoxyphenyl)vinyl]benzofuran as the otherstereoisomer.

¹ H-NMR (CDCl₃) δ:3.84 (3H, s), 6.35 (1H, d, J=14Hz), 6.53 (1H, s), 6.62(1H, d, J=14Hz), 6.9 (2H, d), 7.24 (2H), 7.3 (2H, d), 7.38 (1H)

b) 84 g of a mixture of the two stereoisomers of5-bromo-2-[2-(4-methoxyphenyl)vinyl]benzofuran obtained in the abovestep a) was dissolved in 600 ml of dichloromethane. With ice cooling andstirring, 18.5 ml of acetyl chloride was added to the above solution,followed by dropwise addition of 28.9 ml of titanium tetrachloride. Theresulting reaction solution was poured into ice water, and extractedwith chloroform, and the resulting organic layer was washed with dilutehydrochloric acid and then with water, followed by drying to distill offthe solvent. Thereafter, the resulting residue was suspended in ether tocollect insoluble crystals by filtration, thereby obtaining 76 g of3-acetyl-5-bromo-2-[2-(4-methoxyphenyl)vinyl]benzofuran in the form ofyellow fine needle crystals (the same isomer is formed from both of theE and Z forms).

mp: 163°-165° C.

¹ H-NMR (CDCl₃) δ:2.69 (3H, s), 3.85 (3H, s), 6.95 (2H, d, J=10Hz), 7.4(2H, m), 7.6 (2H, d, J=10Hz), 7.65 (2H, s), 8.08 (1H)

c) A mixture of 20.7 g of3-acetyl-5-bromo-2-[2-(4-methoxyphenyl)vinyl]benzofuran obtained in theabove step b), 6 g of cuprous cyanide and 800 ml ofN-methyl-2-pyrrolidone was stirred at 210° to 220° C. for 8.5 hours in astream of nitrogen. The resulting reaction solution was poured into icewater to remove precipitated materials by filtration, and the resultingfiltrate was extracted with ethyl acetate. After removing insolublematerials by filtration, the resulting organic layer was washed withwater and dried to distill off the solvent. The residue thus obtainedwas subjected to silica gel column chromatography using toluene as aneluant, and the resulting product was washed with methanol. In this way,7.82 g of 3-acetyl-2-[2-(4-methoxyphenyl)vinyl]-5-benzofurancarbonitrilewas obtained in the form of yellow fine crystals.

mp: 190°-191° C.

¹ H-NMR (CDCl₃) δ:2.69 (3H, s), 3.85 (3H, s), 6.98 (2H, d, J=10Hz),7.50-7.80 (6H, m), 8.36 (1H)

d) 7.8 g of3-acetyl-2-[2-(4-methoxyphenyl)vinyl]-5-benzofurancarbonitrile obtainedin the above step c) was dissolved in a solvent mixture of 600 ml oftetrahydrofuran and 500 ml of ethanol. 900 mg of palladium oxide.1H₂O.barium sulfate was added to the above solution, and the mixture wassubjected to catalytic hydrogenation under normal pressure for 3.5hours. After removing the catalyst by filtration, the resulting filtratewas concentrated to dryness. The residue thus obtained was extractedwith ethyl acetate, and the resulting organic layer was washed withwater, and dried to distill off the solvent. Thereafter, the residuethus obtained was washed with methanol to collect precipitated crystalsby filtration, thereby obtaining 5.47 g of3-acetyl-2-[2-(4-methoxyphenyl)ethyl]-5-benzofurancarbonitrile in theform of colorless prism crystals.

mp: 130°-131° C.

¹ H-NMR (CDCl₃) δ: 2.54 (3H, s), 3.04 (2H, m), 3.4 (2H, m), 3.77 (3H,s), 6.85 (2H, d, J=10Hz), 7.05 (2H, d), 7.57 (2H, s), 8.33 (1H)

e) 5.2 g of sodiumhydroxide was dissolved in 30 ml of water, and thesolution was cooled to a temperature of 0° C. or below. With stirring,to this were added dropwise 2.7 ml of bromine and then 40 ml of adioxane solution containing 4.14 g of3-acetyl-2-[2-(4-methoxyphenyl)ethyl]-5-benzofurancarbonitrile obtainedin the above step d). The resulting mixture was stirred at 0° C. for 45minutes with ice cooling for 1 hour. The resulting reaction solution wasmixed with water, adjusted to pH 2 with concentrated hydrochloric acid,and then extracted with chloroform. The resulting organic layer waswashed with water and dried to distill off the solvent. Thereafter, theresulting residue was purified by subjecting it to silica gel columnchromatography using chloroform as an eluant, thereby obtaining 1.44 gof 5-cyano-2-[2-(4-methoxyphenyl)ethyl]-3-benzofurancarboxylic acid.

mp: 205°-208° C. (recrystallization from methanol, fine prism crystals)

¹ H-NMR (CDCl₃) δ:3.13 (2H, m), 3.5 (2H, m), 3.78 (3H, s), 6.83 (2H, d),7.07 (2H, d), 7.56 (2H, s), 8.34 (1H)

f) 1.81 g of5-cyano-2-[2-(4-methoxyphenyl)ethyl]-3-benzofurancarboxyliic acidobtained in the above step e) was added to 5 ml of thionyl chloride, andthe mixture was refluxed under heated for 1 hour. The resulting reactionsolution was concentrated to dryness, and the residue was mixed withethanol and stirred at 50° C. for 30 minutes. Crystals thus precipitatedwere collected by filtration to obtain 1.82 g of ethyl5-cyano-2-(2-(4-methoxyphenyl)ethyl)-3-benzofurancarboxylate.

mp: 135°-139° C. (fine prism crystals)

IR (KBr): 2224, 1695, 1614, 1587, 1515 cm⁻¹

g) 1.78 g of ethyl5-cyano-2-[2-(4-methoxyphenyl)ethyl]-3-benzofurancarboxylate obtained inthe above step f) was treated in the same manner as described in step e)of Reference Example 46 to obtain 2.27 g of the title compound in theform of fine needle crystals.

mp: 182°-183° C.

¹ H-NMR (CDCl₃) δ:1.45 (3H, t, J=8.0Hz), 3.0 (2H, m), 3.4 (2H, m), 4.4(2H, q, J=8.0Hz), 6.7 (2H, d), 7.1 (2H, d), 7.55 (2H), 8.29 (1H)

REFERENCE EXAMPLE 50 Preparation of ethyl[5-cyano-2-(2-(4-hydroxyphenyl)ethyl]-3-benzofuranyl)acetate

a) 128 g of 5-bromo-2-[2-(4-methoxyphenyl)vinyl]benzofuran as a mixtureof two stereoisomers was dissolved in a solvent mixture of 1.3 l oftetrahydrofuran and 0.7 l of ethanol. The resulting solution was mixedwith 3.0 g of platinum dioxide and subjected to 4 hours of catalytichydrogenation undernormal pressure. Thereafter, the catalyst was removedby filtration, the resulting filtrate was concentrated, and crystalsthus precipitated were collected by filtration and washed with ethanol.In this way, 97.08 g of 5-bromo-2-[2-(4-methoxyphenyl)ethyl]benzofuranwas obtained.

mp: 109°-111° C.

¹ H-NMR (CDCl₃) δ: 3.00 (4H, s), 3.77 (3H, s), 6.28 (1H, s), 6.88 (2H,d, J=9.0Hz), 7.08 (2H, d, J=9.0Hz), 7.32 (2H), 7.60 (1H)

b) 97 g of 5-bromo-2-[2-(4-methoxyphenyl)ethyl]benzofuran obtained inthe above step a) was treated in the same manner as described in step b)of Reference Example 49 to obtain 79.9 g of3-acetyl-5-bromo-2-[2-(4-methoxyphenyl)ethyl]benzofuran.

mp: 100°-101° C.

¹ H-NMR (CDCl₃) δ:2.52 (3H, s), 3.05 (2H, m), 3.35 (2H, m), 3.76 (3H,s), 6.80 (2H, d, J=9.0Hz), 7.10 (2H, d, J=9.0Hz), 7.35 (2H, m), 8.05(1H)

c) 79.9 g of 3-acetyl-5-bromo-2-[2-(4-methoxyphenyl)ethyl]benzofuranobtained in the above step b) was treated in the same manner asdescribed in step e) of Reference Example 49 to obtain 64.2 g of5-bromo-2-[2-(4-methoxyphenyl)ethyl]-3-benzofurancarboxylic acid.

¹ H-NMR (DMSO-d₆) δ:3.00 (2H, m), 3.35 (2H, m), 3.69 (3H, s), 6.80 (2H,d, J=8.0Hz), 7.07 (2H, d,-J=8.0Hz), 7.50 (1H, dd), 7.55 (1H, d), 8.00(1H, d)

d) 64.2 g of 5-bromo-2-[2-(4-methoxyphenyl)ethyl]-3-benzofurancarboxylicacid obtained in the above step c) was suspended in 900 ml of ethanol.30 ml of thionyl chloride was added dropwise to the above suspensionwith ice cooling. After refluxing under heating for 5 hours, 50 ml ofthionyl chloride was further added dropwise to the resulting reactionmixture, followed by additional refluxing under heating for 3 hours. Thereaction solution thus obtained was concentrated to dryness, and theresulting residue was mixed with water to collect insoluble materials byfiltration. The thus collected insoluble materials were dissolved inethyl acetate and washed with a saturated sodium bicarbonate aqueoussolution, water, a saturated sodium chloride aqueous solution in thatorder, followed by drying to distill off the solvent. Thereafter, theresulting residue was suspended in ethanol and then collected byfiltration to obtain 59.23 g of ethyl5-bromo-2-[2-(4-methoxyphenyl)ethyl]-3-benzofurancarboxylate.

mp: 73°-75° C.

¹ H-NMR (CDCl₃) δ:1.43 (3H, t, J=8.9Hz), 3.10 (2H, m), 3.40 (2H, m),3.77 (3H, s), 4.40 (2H, q, J=8.9Hz), 6.80 (2H, d), 7.2 (2H, d), 7.33(2H, m), 8.10 (1H)

e) 35.5 g of ethyl5-bromo-2-[2-(4-methoxyphenyl)ethyl]-3-benzofurancarboxylate obtained inthe above step d) was dissolved in 400 ml of tetrahydrofuran, followedby the gradual addition of 3.5 g of lithium-aluminum hydride and,subsequently, 1 hour of stirring at room temperature. The resultingreaction solution was poured into water, adjusted to pH 2 withhydrochloric acid and then extracted with benzene. Thereafter, theresulting organic layer was washed with water, and then concentrated todryness to obtain 30 g of5-bromo-3-hydroxymethyl-2-[2-(4-methoxyphenyl)ethyl]benzofuran in theform of crystals.

mp: 65°-75° C.

¹ H-NMR (CDCl₃) δ:2.95 (4H, s), 3.69 (3H, s), 4.33 (2H, s), 6.77 (2H,d), 6.90 (2H, d), 7.26 (2H, m), 7.65 (1H)

f) 30 g of5-bromo-3-hydroxymethyl-2-[2-(4-methoxyphenyl)ethyl]benzofuran obtainedin the above step e) was suspended in 150 ml of diethyl ether. To thiswas added 12 drops of pyridine. With ice cooling, 12 ml of thionylchloride was further added dropwise. The reaction mixture thus preparedwas stirred for 1 hour at room temperature. The resulting reactionsolution was poured into ice water, and extracted with diethyl ether.Thereafter, the resulting organic layer was washed with water and thenWith saturated sodium bicarbonate aqueous solution, followed byconcentration to dryness, thereby obtaining 28.3 g of5-bromo-3-chloromethyl-2-[2-(4-methoxyphenyl)ethyl]benzofuran.

mp: 70°-75° C.

¹ H-NMR (CDCl₃) δ: 3.00 (4H, s), 3.76 (3H, s), 4.38 (2H, s), 6.82 (2H,d), 6.97 (2H, d), 7.31 (2H), 7.68 (1H)

g) To 75 ml of acetonitrile were added 10.82 g of5-bromo-3-chloromethyl-2-[2-(4-methoxyphenyl)ethyl]benzofuran obtainedin the above step f), 3.7 g of potassium cyanide and 0.6 g of18-crown-6-ether. The thus prepared mixture was refluxed under heatingfor 2.5 hours. The reaction solution thus obtained was mixed with water,and extracted with benzene, and the resulting organic layer was washedwith water, and dried to distill off the solvent. Thereafter, theresulting residue was purified by subjecting it to silica gel columnchromatography using a toluene/n-hexane mixture as an eluant, therebyobtaining 9.17 g of5-bromo-3-cyanomethyl-2-[2-(4-methoxyphenyl)ethyl]benzofuran.

¹ H-NMR (CDCl₃) δ:2.95 (4H, s), 3.20 (2H, s), 3.73 (3H, s), 6.80 (2H,d), 6.90 (2H, d), 7.33 (2H), 7.61 (1H)

h) 9.17 g of5-bromo-3-cyanomethyl-2-[2-(4-methoxyphenyl)ethyl]benzofuran obtained inthe above step g) was added to a mixture solution of 100 ml of ethanoland 5 ml of concentrated sulfuric acid, and the resulting mixture wasrefluxed under heating for 18 hours. The resulting reaction solution waspoured into water and extracted with ethyl acetate, Thereafter, theresulting organic layer was washed with water, a saturated sodiumbicarbonate aqueous solution and water in that order, followed by dryingto distill off the solvent. In this way, 8.96 g of ethyl[5-bromo-2-[2-(4-methoxyphenyl)ethyl]-3-benzofuranyl]acetate wasobtained.

¹ H-NMR (CDCl₃) δ:1.21 (3H, t, J=7.0Hz), 2.96 (4H, s), 3.34 (2H, s),3.74 (3H, s), 4.10 (2H, q, J=7.0Hz), 6.80 (2H, d, J=9Hz), 7.00 (2H, d,J=7.0Hz), 7.28 (2H), 7.59 (1H)

i) 8.2 g of ethyl[5-bromo-2-[2-(4-methoxyphenyl)ethyl]3-benzofuranyl]acetate obtained inthe above step h) was treated in the same manner as described in step c)of Reference Example 49 to obtain 4.5 g of ethyl[5-cyano-2-[2-(4-methoxyphenyl)ethyl]-3-benzofuranyl]acetate in the formof colorless needle crystals.

mp: 85°-86° C.

¹ H-NMR (CDCl₃) δ:1.23 (3H, t, J=7.0Hz), 3.01 (4H, s), 3.40 (2H, s),3.75 (3H, s), 4.11 (2H, q, J=7.0Hz), 6.80 (2H, d, J=9Hz), 7.00 (2H, d,J=7.0Hz), 7.47 (2H), 7.81 (1H)

j) 4.45 g of ethyl[5-cyano-2-[2-(4-methoxyphenyl)ethyl]-3-benzofuranyl]acetate obtained inthe above step i) was treated in the same manner as described in step e)of Reference Example 46 to obtain 2.98 g of the title compound in theform of colorless crystals.

mp: 134°-136° C.

¹ -NMR (CDCl₃) δ:1.22 (3H, t, J=7.0Hz), 2.98 (4H, s), 3.39 (2H, s), 4.12(2H, q, J=7.0Hz), 6.74 (2H, q, J=9.0Hz), 6.91 (2H, d, J=7.0Hz), 7.48(2H), 7.80 (1H)

REFERENCE EXAMPLE 51 Preparation of ethyl3-[2-[2-(5-cyanobenzo[b]thien-2-yl)ethyl]-4-ethoxy-5-hydroxyphenyl]propionate

a) 20.0 g of Ferulic acid was dissolved in 250 ml of methanol andsubjected to catalytic reduction under normal pressure for 3 hours inthe presence of 10% palladium carbon catalyst (50% wet type). Afterremoving the catalyst by filtration, the resulting filtrate wasconcentrated to collect precipitated crystals by filtration, therebyobtaining 19.3 g of 3-(4-hydroxy-3-methoxyphenyl)propionic acid.

mp: 87°-89° C.

¹ H-NMR (CDCl₃) δ:2.5-3.0 (4H, m), 3.85 (3H, s), 6.5-6.9 (3H, m)

b) 19.3 g of 3-(4-hydroxy-3-methoxyphenyl)propionic acid obtained in theabove step a) was dissolved in 300 ml of ethanol. After adding 2.0 ml ofconcentrated sulfuric acid, the resulting mixture was refluxed underheating for 2 hours. The resulting reaction solution was concentratedunder a reduced pressure, extracted with chloroform, washed with water,and then dried. By distilling off the solvent, 23.0 of ethyl3-(4-hydroxy-3-methoxyphenyl)propionate was obtained in an oily form.

¹ H-NMR (CDCl₃) δ:1.23 (3H, t, J=7.2Hz), 2.4-3.0 (4H, m), 3.85 (3H, s),4.12 (2H, q, J=7.12Hz), 6.6-6.9 (3H, m)

c) 10.0 g of ethyl 3-(4-hydroxy-3-methoxyphenyl)propionate obtained inthe above step b) was dissolved in 300 ml of tetrahydrofuran. 1.96 g of60% sodium hydride was subsequently added. The thus prepared mixture wasstirred at 50° C. for 30 minutes. To this was added dropwise 7.17 g ofethyl bromide. After refluxing under heating for 6 hours, the resultingreaction solution was poured into water, extracted with chloroform,washed with water, and then concentrated under a reduced pressure.Thereafter, the resulting residue was subjected to silica gel columnchromatography using chloroform as an eluant, thereby obtaining 5.6 g ofoily ethyl 3-(4-ethoxy-3-methoxyphenyl)propionate.

¹ H-NMR (CDCl₃) δ:1.23 (3H, t, J=7.2Hz), 1.43 (3H, t, J=7.1Hz), 2.4-3.0(4H, m), 3.85 (3H, s), 4.06 (2H, q, J=7.1Hz), 4.11 (2H, q, J=7.12Hz),6.7-6.9 (3H, m)

d) 9.3 g of ethyl 3-(4-ethoxy-3-methoxyphenyl)propionate obtained in theabove step c) was dissolved in 10 ml of acetic acid, followed by adding7.4 g of chloromethyl methyl ether and subsequently stirring at roomtemperature for 22 hours. The resulting reaction solution was pouredinto ice water, extracted with ethyl acetate, washed with water and thendried to distill off the solvent. The residue thus obtained wasdissolved-in 10 ml of xylene, and the solution was mixed with 8.54 g oftriphenylphosphine. The resulting mixture was stirred at roomtemperature for 18 hours and then at 70° to 80° C. for 5 hours. Aftercooling, xylene was removed by decantation, and the remaining portionwas solidified by adding n-hexane to obtain 6.0 g of crude[5-ethoxy-2-(2-ethoxycarbonylethyl)-4-methoxyphenyl]methyltriphenylphosphoniumchloride.

e) To a mixture solution of 50 ml of tetrahydrofuran and 50 ml ofethanol were dissolved 1.5 g of 5-cyanobenzo[b]thiophene-2-carbaldehydeand 6.34 g of crude[5-ethoxy-2-(2-ethoxycarbonylethyl)-4-methoxyphenyl]methyltriphenylphosphoniumchloride obtained in the above step d). 1.83 g of1,8-diazabicyclo[5.4.0]-7-undecene was added thereto, followed bystirring at room temperature for 18 hours. The resulting reactionsolution was concentrated under a reduced pressure, and the residue wasdissolved in a mixture solution of 20 ml of tetrahydrofuran and 20 ml ofethanol. The thus prepared solution was mixed with 1.70 g of 10%palladium carbon catalyst (50% wet type) and subjected to catalytichydrogenation under normal pressure until hydrogen absorption wascompleted. After removing the catalyst by filtration, the solvent wasdistilled off. By subjecting the resulting residue to silica gel columnchromatography, 1.2 g of oily ethyl3-[2-[2-(5-cyanobenzo[b]thien-2-yl)ethyl]-4-ethoxy-5-methoxyphenyl]propionatewas obtained.

¹ H-NMR (CDCl₃) δ:1.24 (3H, t, J=7.1Hz), 1.36 (3H, t, J=7.0Hz), 2.4-3.3(8H, m), 3.84 (3H, s), 3.98 (2H, q, J=7.0Hz), 4.13 (2H, q, J=7.1Hz),6.64 (1H, s), 6.70(1H, s), 7.04 (1H, s), 7.46 (1H, dd, J=8.4 and 1.5Hz),7.83(1H, d, J=8.4Hz), 7.96 (1H, s)

f) 2.1 g of ethyl3-[2-[2-(5-cyanobenzo[b]thien-2-yl)ethyl]-4-ethoxy-5-methoxyphenyl]propionateobtained in the above step e) was dissolved in 20 ml of γ-collidine,followed by adding 7.94 g of lithium iodide and subsequently refluxingunder heating for 18 hours. The resulting reaction solution was pouredinto water, extracted with chloroform, washed with water, and thendried. After distilling off the solvent, the residue thus obtained wasdissolved in 100 ml of ethanol, mixed with 0.3 ml of concentratedsulfuric acid, and then refluxed under heating for 1 hour. Afterdistilling off the solvent under a reduced pressure, the resultingreaction solution was diluted with chloroform, washed with water, andthen dried to distill off the solvent. Thereafter, the resulting residuewas subjected to purification by subjecting it to silica gel columnchromatography using chloroform as an eluant. In this way, 2.0 g of thetitle compound was obtained in an oily form.

¹ H-NMR (CDCl₃) δ:1.24 (3H, t, J=7.1Hz), 1.35 (3H, t, J=7.0Hz), 2.4-3.3(8H, m), 3.98 (2H, q, J=7.0Hz), 4.13 (2H, q, J=7.1Hz), 6.60(1H, s), 6.75(1H, s), 7.44 (1H, dd, J =8.4 and 1.5Hz), 7.82 (1H, d, J=8.4Hz), 7.96(1H, s), 7.94 (1H, s)

REFERENCE EXAMPLE 52 Preparation ofethyl-3-[3-[2-(5-cyanobenzo[b]thien-2-yl)ethyl]-5-hydroxyphenyl]propionate

a) 33.5 g of 6-methoxy-2-tetralone was dissolved in 27.6 ml of ethanol,followed by subsequently adding 37.8 ml of ethyl orthoformate and onedrop of concentrated sulfuric acid. The mixture2thus prepared wasstirred at 100° C. for 4 hours. After distilling off the solvent under areduced pressure, the resulting residue was subjected to silica gelcolumn chromatography using chloroform as an eluant. Fractions ofinterest were pooled and concentrated to collect precipitated crystals,thereby obtaining 5.82 g of 3,4-dihydro-2-ethoxy-6-methoxynaphthalene.

¹ H-NMR (CDCl₃) δ:1.37 (3H, t, J=7.0Hz), 2.20-3.00 (4H, m), 3.79 (3H,s), 3.84 (2H, q, J=7.0Hz), 5.48 (1H, s), 6.60-7.00 (3H, m)

b) 5.8 g of 3,4-dihydro-2-ethoxy-6-methoxy-naphthalene obtained in theabove step a) was dissolved in a mixture solution of 90 ml of ethanoland 10 ml of dichloromethane. With stirring at a cooled temperature of-20° C., ozone was bubbled into the solution prepared above to effectoxidation. 10 ml of dimethylsulfide was added dropwise and gradually tothe resulting reaction solution at the same temperature, followed bystirring at room temperature for 30 minutes. After distilling off thesolvent under a reduced pressure, the resulting residue was dissolved in100 ml of a tetrahydrofuran/ethanol mixture (1:1). To this were added12.5 g of (5-cyanobenzo[b]thien-2-yl)methyltriphenylphosphonium chlorideand 4.46 ml of 1,8-diazabicyclo[5.4.0]-7-undecene in that order,followed by 5 hours of stirring. The resulting reaction solution wasconcentrated under a reduced pressure, and the residue thus obtained waspurified by subjecting it to silica gel column chromatography usingchloroform as an eluant. The thus purified product was dissolved in 60ml of ethanol/tetrahydrofuran mixture (1:1), and the resulting solutionwas mixed with 3.9 g of 10% palladium carbon catalyst (50% wet type). Bysubjecting the thus prepared mixture to catalytic hydrogenation undernormal pressure for 3 hours, 2.75 g of ethyl3-[2-[2-(5-cyanobenzo[b]thien-2-yl)ethyl]-5-methoxyphenyl]propionate wasobtained.

¹ H-NMR (CDCl₃) δ:1.22 (3H, t, J=7.2Hz), 2.2-3.4 (8H, m), 3.76 (3H, s),4.16 (2H, q, J=7.2Hz), 6.60-7.30 (4H, m), 7.48 (1H, d, J=8.2Hz), 7.85(1H, d, J=8.2Hz), 7.99 (1H, s)

c) 2.75 g of ethyl3-[2-[2-(5-cyanobenzo[b]-thien-2-yl)ethyl]-5-methoxyphenyl)propionateobtained in the above step b) was treated in the same manner asdescribed in step e) of Reference Example 46 to obtain 2.3 g of thetitle compound in an oily form.

¹ H-NMR (CDCl₃) δ:1.23 (3H, t, J=7.1Hz), 2.4-3.34 (8H, m), 4.13 (2H, q,J=7.2Hz), 5.60 (1H, s), 6.50-7.20 (3H, m), 7.25 (1H, s), 7.44 (1H, d,J=8.3Hz), 7.82 (1H, d, J=8.3Hz), 7.92 (1H, s)

REFERENCE EXAMPLE 53 Preparation of ethyl2-(5-cyanobenzo[b]thien-2-yl)-3-(4-hydroxyphenyl]propionate

a) 0.5 g of 5-bromo-2-hydroxymethylbenzo[b]thiophene was dissolved in 20ml of dichloromethane, followed by the addition of 230 mg of phosphorustribromide. After 1 hour of stirring at room temperature, the resultingreaction solution was mixed with water, washed with a saturated sodiumbicarbonate aqueous solution, and then dried to distill off the solvent.The resulting residue was dissolved in a mixture solvent of 10 ml ofacetonitrile and 3 ml of dimethylsulfoxide, followed by the addition of300 mg of cuprous cyanide and subsequently refluxing under heating for 2hours. After cooling, toluene was added to the reaction solution toremove insoluble materials by filtration, and the resulting filtrate waswashed with water, dried, and concentrated. By collecting precipitatedcrystals by filtration, 200 mg of 5-bromo-2-cyanomethylbenzo[b]thiophenewas obtained.

mp: 94°-96° C.

¹ H-NMR (CDCl₃) δ:3.98 (2H, s), 7.25 (1H, s), 7.42 (1H, dd, J=8.5 and1.8Hz), 7.65 (1H, d, J=8.5Hz), 7.90 (1H, d, J=1.8Hz)

b) 12.0 g of 5-bromo-2-cyanomethylbenzo[b]thiophene obtained in theabove step a) was dissolved in 80 ml of ethanol, followed by theaddition of 1.0 ml of water and 7 ml of concentrated sulfuric acid.After refluxing under heating for 7 hours, the resulting reactionsolution was mixed with 40 ml of ethanol, 15 ml of concentrated sulfuricacid and 0.5 ml of water, and the mixture was refluxed under heating for2 hours. After cooling, the resulting reaction solution was mixed withwater, and extracted with an equal volume mixture of toluene and ethylacetate. The resulting organic layer was washed with water and saturatedsodium bicarbonate aqueous solution in that order, and then dried todistill off the solvent. Thereafter, the resulting residue was subjectedto silica gel column chromatography to obtain 8.0 g of ethyl2-(5-bromobenzo[b]thien-2-yl)acetate.

mp: 56°-57° C.

¹ H-NMR (CDCl₃) δ:1.28 (3H, t, J=7.0Hz), 3.88 (2H, s), 4.23 (2H, q,J=7.0Hz), 7.11 (1H, s), 7.38 (1H, dd, J=8.3 and 1.8Hz), 7.68 (1H, d,J=8.3Hz), 7.82 (1H, d, J=1.8Hz)

c) 800 mg of ethyl 2-(5-bromobenzo[b]thien-2-yl)acetate obtained in theabove step b) and 965 mg of diethyl carbonate were dissolved in 4 ml ofN,N-dimethylformamide. With heating in an oil bath of 120° to 130° C.,162 mg of sodium hydride (60%) was added to the above solution. After 10minutes of stirring at the same temperature, 30 mg of sodium hydride(60%) was added to the reaction solution, and the stirring was continuedfor another 10 minutes. The resulting reaction solution was diluted withan equal volume mixture of toluene and ethyl acetate, washed with dilutehydrochloric acid and water in that order, and then dried. Afterdistilling off the solvent, the resulting residue was purified bysubjecting it to silica gel column chromatography using a toluene/ethylacetate mixture as an eluant. In this way, 600 mg of ethyl2-(5-bromobenzo[b]thien-2-yl)-2-ethoxycarbonylacetate.

¹ H-NMR (CDCl₃) δ:1.28 (6H, t, J=7.0Hz), 4.25 (4H, q, J=7.0Hz), 4.95(1H, s), 7.17 (1H, s), 7.35 (1H, dd, J=8.3 and 2.1Hz), 7.62 (1H, d,J=8.3Hz), 7.83 (1H, d, J=2.1Hz)

d) 6.2 g of ethyl 2-(5-bromobenzo[b]thien-2-yl)-2-ethoxycarbonylacetateobtained in the above step c) and 5.2 g of 4-methoxybenzyl chloride weredissolved in 30 ml of N,N-dimethylformamide. At room temperature, 1.34 gof sodium hydride (60%) was added to the above solution, and the mixturewas stirred for 3 hours. With ice cooling, the resulting reactionsolution was mixed with 10% citric acid aqueous solution, extracted withtoluene, washed with water, and then dried. After distilling off thesolvent, the resulting residue was subjected to silica gel columnchromatography using toluene as an elution solvent to obtain 8.2 g ofethyl2-(5-bromobenzo[b]thien-2-yl)-2-ethoxycarbonyl-3-(4-methoxyphenyl)propionate.

mp: 58°-60° C.

¹ H-NMR (CDCl₃) δ:1.22 (6H, t, J=7.0Hz), 3.65 (5H, s), 4.30 (4H, q,J=7.0Hz), 6.60 (2H, d, J=8.5Hz), 6.79 (2H, d, J=8.5Hz), 7.31 (1H, s),7.35 (1H, dd, J=8.8 and 1.8Hz), 7.60 (1H, d, J=8.8Hz), 7.82 (1H, d,J=1.8Hz)

e) 3.0 g of ethyl2-(5-bromobenzo[b]thien-2-yl)-2-ethoxycarbonyl-3-(4-methoxyphenyl)propionateobtained in the above step d) was dissolved in 25 ml of ethanol. 0.91 gof potassium hydroxide dissolved in 2.5 ml of water was added to theabove solution, and the mixture was stirred at room temperature for 4days. With ice cooling, the thus obtained reaction solution was mixedwith dilute hydrochloric acid and extracted with ethyl acetate, and theresulting organic layer was washed with water, and dried to distill offthe solvent. The resulting residue was dissolved in 60 ml of ethanol,mixed with 4 ml of concentrated sulfuric acid, and then refluxed underheating for 1 hour. After ice cooling, the resulting reaction solutionwas washed with saturated sodium bicarbonate aqueous solution, and thenwith water, followed by drying to distill off the solvent. Thereafter,the resulting residue was purified by subjecting it to silica gel columnchromatography to obtain 1.6 g of ethyl2-(5-bromobenzo[b]thien-2-yl)-3-(4-methoxyphenyl)propionate.

mp: 62°-65° C.

¹ H-NMR (CDCl₃) δ: 1.15 (3H, t, J=7.0Hz), 3.08 (1H, dd, J =13.5 and7.3Hz), 3.37 (1H, dd, J=13.5 and 7.3Hz), 3.71 (3H, s), 4.10 (2H, q,J=7.0Hz), 4.14 (1H, t, J=7.3Hz), 6.75 (2H, d, J=8.7Hz), 6.83 (1H, s),7.05 (2H, d, J=8.7Hz), 7.30 (1H, dd, J=8.8 and 2.3Hz), 7.57 (1H, d,J=8.8Hz), 7.74 (1H, d, J=2.3Hz)

f) 1.6 g of ethyl2-(5-bromobenzo[b]thien-2-yl)-3-(4-methoxyphenyl)propionate obtained inthe above step e) was treated in the same manner as described in step c)of Reference Example 49 to obtain 1.0 g of ethyl2-(5-cyanobenzo[b]thien-2-yl)-3-(4-methoxyphenyl)propionate.

mp: 93°-96° C.

¹ H-NMR (CDCl₃) δ:1.17 (3H, t, J=7.0Hz), 3.09 (1H, dd, J =14.0 and8.0Hz), 3.39 (1H, dd, J=14.0 and 8.0Hz), 3.73 (3H, s), 4.12 (2H, q,J=7.0Hz), 4.16 (1H, t), 6.75 (2H, d, J=8.8Hz), 7.05 (2H, d, J=8.8Hz),7.13 (1H, s), 7.43 (1H, dd, J=8.3 and 1.3Hz), 7.81 (1H, d, J=8.3Hz),7.92 (1H, br)

g) 3.3 g of ethyl2-(5-cyanobenzo[b]thien-2-yl)-3-(4-methoxyphenyl)propionate obtained inthe above step f) was treated in the same manner as described in step e)of Reference Example 46 to obtain 2.8 g of the title compound.

mp: 146°-147° C.

¹ H-NMR (CDCl₃) δ:1.19 (3H, t, J=7.0Hz), 3.09 (1H, dd, J =13.5 and7.5Hz), 3.38 (1H, dd, J=13.5 and 7.5Hz), 4.13 (2H, q, J=7.0Hz), 4.18(1H, t), 6.70 (2H, d, J=8.5Hz), 7.00 (2H, d, J=8.5Hz), 7.15 (1H, s),7.47 (1H, dd, J=8.3 and 1.3Hz), 7.85 (1H, d, J=8.3Hz), 7.95 (1H, br)

REFERENCE EXAMPLE 54 Preparation of ethyl2-[4-[((2S)-1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxy]phenyl]-3-(5-cyano-2-benzofuranyl)propionate

1.04 g of diethyl azodicarboxylate was added to 300 ml of atetrahydrofuran solution containing 1 g of ethyl3-(5-cyano-2-benzofuranyl )-2-(4-hydroxyphenyl)propionate, 1.2 g of(2S)-1-tert-butoxycarbonyl-2-pyrrolidinemethanol and 1.56 g oftriphenylphosphine, and the thus prepared mixture was stirred at roomtemperature for 18 hours. To the resulting reaction solution were added0.6 g of (2S)-1-tert-butoxycarbonyl-2-pyrrolidinemethanol, 0.78 g oftriphenylphosphine and 0.52 g of diethyl azodicarboxylate, followed byadditional stirring at room temperature for 18 hours. Thereafter, thereaction solution thus obtained was concentrated to dryness, and theresulting residue was purified by subjecting it to silica gel columnchromatography using a toluene/ethyl acetate mixture as an eluant. Inthis way, 790 mg of the title compound was obtained in the form ofcolorless oil.

¹ H-NMR (CDCl₃) δ:1.15 (3H, t), 1.46 (9H, s), 1.98 (4H, br), 3.0-4.2(8H, m), 4.1 (2H), 6.37 (1H, s), 6.9 (2H, d), 7.2 (2H, d), 7.45 (2H),7.76 (1H, s)

The following compounds of Reference Examples 55 to 61 were prepared inaccordance with the procedure of Reference Example 54.

REFERENCE EXAMPLE 55 ethyl3-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-benzofuranyl)propionate

¹ H-NMR (CDCl₃) δ:1.16 (3H, t, J=7Hz), 1.46 (9H, s), 2.0-2.2 (2H, m),2.8-3.2 (2H, m), 3.5-3.7 (4H, m), 4.10 (2H, q, J=7Hz), 4.5-4.7 (1H, m),4.9 (1H, m), 6.49 (1H, s), 6.82 (2H, d, J=9Hz), 7.23 (2H, d, J=9Hz),7.47 (2H, s), 7.80 (1H, s)

REFERENCE EXAMPLE 56 ethyl5-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]-2-[-2-(5-cyano-2-benzofuranyl)ethyl]benzoate

¹ H-NMR (CDCl₃) δ:1.38 (3H, t, J=7.0Hz), 1.48 (9H, s), 2.00-2.30 (2H,m), 2.96-3.76 (8H, m), 4.36 (2H, q), 4.90 (1H, br), 6.44 (1H, s), 6.93(1H, dd, J=8.8 and 2.7Hz), 7.15 (1H, d, J=8.8Hz), 7.48 (1H, d, J=2.7Hz),7.52 (2H, s), 7.80 (1H, s)

REFERENCE EXAMPLE 57 ethyl2-[5-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]-2-[2-(5-cyano-2-benzofuranyl)ethyl]phenyl]acetate

¹ H-NMR (CDCl₃) δ:1.25 (3H, t, J=7.0Hz), 1.47 (9H, s), 1.90-2.30 (2H,m), 3.04 (4H, s), 3.36-3.70 (6H, m), 4.16 (2H, q, J=7.0Hz), 4.90-5.12(1H, br), 6.42 (1H, s), 6.60-6.80 (2H, m), 7.08 (1H, d, J=7.6Hz), 7.48(2H, s), 7.77 (1H, d, J=0.87Hz)

REFERENCE EXAMPLE 58 ethyl2-[2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]ethyl]-5-cyano-3-benzofurancarboxylate

¹ H-NMR (CDCl₃) δ:1.46 (12H, m), 2.05 (2H, m), 2.95 (2H, m), 3.5 (6H,m), 4.4(2H, q), 4.80 (1H, br), 6.82 (2H, d), 7.08 (2H, d), 7.55 (2H),8.30 (1H)

REFERENCE EXAMPLE 59 ethyl3-[5-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]2-[2-(5-cyanobenzo[b]thien-2-yl)ethyl]-4-ethoxyphenyl]propionate

¹ H-NMR (CDCl₃) δ:1.23 (3H, t, J=7.2Hz), 1.33 (3H, t, J=7.0Hz), 1,47(9H, s), 2.00-3.30 (10H, m), 3.4-3.7 (4H, m), 3.94 (2H, q, J=7.0Hz),4.12 (2H, q, J=7.2Hz), 4.70-5.00 (1H, br), 6.67 (1H, s), 6.72 (1H, s),7.05 (1H, s), 7.46 (1H, dd, J=8.4 and 1.6Hz), 7.84 (1H, d, J=8.4Hz),7.95 (1H, d, J=1.6Hz)

REFERENCE EXAMPLE 60 ethyl3-[5-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]-2-[2-(5-cyanobenzo[b]thien-2-yl)ethyl]phenyl]propionate

mp: 117°-119° C.

¹ H-NMR (CDCl₃) δ:1.24 (3H, t, J=7.1Hz), 1.47 (9H, s), 1.6-3.6 (14H, m),4.13 (2H, q, J=7.1Hz), 4.6-4.9 (1H, m), 6.50-7.20 (4H, m), 7.45 (1H, d,J=8.5Hz), 7.83 (1H, d, J =8.5Hz), 7.95 (1H, s)

REFERENCE EXAMPLE 61 ethyl3-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-2-(5-cyanobenzo[b]thien-2-yl)propionate

¹ H-NMR (CDCl₁) δ:1.16 (3H, t, J=7.0Hz), 1.45 (9H, s), 3.08 (1H, dd,J=13.7 and 7.4Hz), 3.30-3.70 (5H, m), 4.11 (2H, q, J=7.0Hz), 4.00-4.30(1H), 6.72 (2H, d, J=8.3Hz), 7.07 (2H, d, J=8.3Hz), 7.10 (1H, s), 7.40(1H, dd, J=8.3 and 1.3Hz), 7.77 (1H, d, J=8.3Hz), 7.89 (1H, br)

REFERENCE EXAMPLE 62 Preparation of methyl3-[5-cyano-2-benzofuranyl]-2-[4-[(tetrahydro-3-furanyl)oxy]phenyl]propionate

In 30 ml of tetrahydrofuran were dissolved 3 g of3-hydroxytetrahydrofuran, 6.5 g of methyl2-(4-hydroxyphenyl)-2-oxoacetate and 9 g of triphenylphosphine. The thusprepared solution was mixed with 6.5 g of diethyl azodicarboxylate, andthe mixture was stirred for 2 hours. After distilling off the solvent,the resulting residue was purified by subjecting it to silica gel columnchromatography using dichloromethane as an eluant, thereby obtaining 7.5g of methyl 2-[4-[(tetrahydro-3-furanyl)oxy]phenyl]-2-oxoacetate in anoily form.

In a mixture solvent of 30 ml of tetrahydrofuran and 50 ml of methanolwere dissolved 2.2 g of methyl2-[4-[(tetrahydro-3-furanyl)oxy]phenyl]-2-oxoacetate obtained above and3.6 g of (5-cyano-2-benzofuranyl)methyltriphenylphosphonium chloride.With ice cooling, to the thus prepared solution was added 1.5 ml of1,8-diazabicyclo[5.4.0]-7-undecene. After 18 hours of stirring at roomtemperature, the reaction solution was concentrated to dryness, and theresulting residue was purified by subjecting it to silica gel columnchromatography using a chloroform/acetone mixture as an eluant, therebyobtaining methyl3-(5-cyano-2-benzofuranyl)-2-[4-[(tetrahydro-3-furanyl)oxy]phenyl]acrylateas a mixture of E and Z forms. The acrylic acid derivative thus obtainedwas dissolved in 80 ml of methanol, mixed with 4 g of palladiumoxide.1H₂ O.barium sulfate and then subjected to catalytic hydrogenationunder normal pressure. Thereafter, the catalyst was removed byfiltration, the resulting filtrate was concentrated to dryness, and thethus obtained residue was purified by subjecting it to silica gel columnchromatography using a chloroform/acetone mixture as an eluant. In thisway, 2.5 g of the title compound was obtained.

¹ H-NMR (CDCl₃) δ:2.0-2.3 (2H, m), 3.2 (1H, dd), 3.6 (1H, dd), 3.65 (3H,s), 3.97 (2H, d), 3.8-4.2 (1H, m), 4.8-5.0 (1H, m), 6.40 (1H, s), 6.8(2H, d), 7.25 (2H, d), 7.5 (2H, s), 7.79 (1H, s)

REFERENCE EXAMPLE 63 Preparation of methyl3-(5-cyano-2-indolyl)-2-[4-[((3R)-tetrahydro-3-furanyl)oxy]phenyl]propionate

In 30 ml of tetrahydrofuran were dissolved 3.0 g of(S)-(+)-3-hydroxytetrahydrofuran, 6.6 g of methyl2-(4-hydroxyphenyl)-2-oxoacetate and 8.90 g of triphenylphosphine. Thethus prepared solution was mixed with 6.0 g of diethyl azodicarboxylate,and the mixture was stirred for 2 hours. After distilling off thesolvent, the resulting residue was purified by subjecting it to silicagel column chromatography using chloroform as an eluant, therebyobtaining 4.60 g of methyl2-[4-[((3R)-tetrahydro-3-furanyl)oxy]phenyl]-2-oxoacetate in an oilyform.

In a mixture solvent of 30 ml of tetrahydrofuran and 30 ml of methanolwere dissolved 1.70 g of methyl2-[4-[((3R)-tetrahydro-3-furanyl)oxy]phenyl]-2-oxoacetate obtained aboveand 3.0 g of (5-cyano-2-indolyl)methyltriphenylphosphonium bromide. Tothe thus prepared solution was added 2.1 ml of1,8-diazabicyclo[5.4.0]-7-undecene during stirring under ice cooling,followed by stirring at room temperature for 2 hours. After distillingoff the solvent, the resulting residue was purified by subjecting it tosilica gel column chromatography using a chloroform/acetone mixture asan elution solvent, thereby obtaining methyl3-(5-cyano-2-indolyl)-2-[4-[((3R)-tetrahydro-3-furanyl)oxy]phenyl]acrylateas a mixture of E and Z forms. The E/Z mixture thus obtained wasdissolved in 50 ml of methanol, mixed with 4.0 g of palladium oxide.1H₂O.barium sulfate and then subjected to catalytic hydrogenation undernormal pressure for 3 hours. Thereafter, the catalyst was removed byfiltration, solvent in the resulting filtrate was distilled off, and thethus obtained residue was purified by subjecting it to silica gel columnchromatography using a chloroform/acetone mixture as an elution solvent.In this way, 1.50 g of the title compound was obtained as a viscous oilymaterial.

¹ H-NMR (CDCl₃) δ:3.10 (1H, dd), 3.60 (3H, s), 3.78-4.10 (5H, m),4.75-5.00 (1H, m), 6.25 (1H, br), 6.80 (2H, d), 7.20 (2H, d), 7.30-7.90(3H, m), 10.00 (1H, s)

REFERENCE EXAMPLE 64 Preparation of methyl3-(5-cyano-2-indolyl)-2-[4-[((3S)-tetrahydro-3-furanyl)oxy]phenyl]propionate

a) In 80 ml of tetrahydrofuran were dissolved 5.0 g of(S)-(+)-3-hydroxytetrahydrofuran, 3.3 g of formic acid and 17.0 g oftriphenylphosphine. With ice cooling and with stirring, 12.0 g ofdiethyl azodicarboxylate was added dropwise to the above solution. Afterstirring at room temperature for 2 hours, the solvent was distilled off,and the resulting residue was purified by subjecting it to silica gelcolumn chromatography using chloroform as an elution solvent, therebyobtaining (S)-(+)-tetrahydro-3-furanyl formate which was subsequentlydissolved in 50 ml of ethanol. With stirring, 5.0 g of sodium hydroxidedissolved in 5 ml of water was added to the above ester solution,followed by stirring for 3 hours. After distilling off the solvent, theresulting residue was dissolved in diethyl ether, and insolublematerials were removed by filtration. By distilling off the solvent,4.50 g of crude (R)-(-)-3-hydroxytetrahydrofuran was obtained.

b) The crude (R)-(-)-3-hydroxytetrahydrofuran obtained in the above stepa) was treated in the same manner as described in Reference Example 63to obtain 1.50 g of the title compound as a viscous oily material.

¹ H-NMR (CDCl₃) δ:3.15 (1H, dd), 3.65 (3H, s), 3.80-4.20 (5H, m),4.80-5.05 (1H, m), 6.30 (1H, br), 6.82 (2H, d), 7.22 (2H, d), 7.30-7.90(3H, m), 9.30 (1H, br)

REFERENCE EXAMPLE 65 Preparation of methyl3-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-2-(5-cyano-2-benzofuranyl)propionate

a) 21.0 g of 2-acetyl-5-benzofurancarbonitrile was dissolved in 300 mlof dichloromethane. With stirring at a temperature of -10° C., 30 ml ofdichloromethane solution containing 18.2 g of bromine was added dropwiseto the above solution. After gradually warming up to ice-coldtemperature, the resulting reaction solution was mixed with chloroformand washed with 10% sodium thiosulfate aqueous solution. After dryingthe organic layer and subsequent concentration to dryness, the resultingresidue was recrystallized from a benzene/n-hexane mixture to obtain21.0 g of 2-(2-bromo-1-oxoethyl)-5-benzofurancarbonitrile in the form ofcolorless crystals.

mp: 156°-158° C.

IR (KBr): 2228, 1696, 1616, 1564, 1290, 1166, 1122 cm⁻¹

¹ H-NMR (CDCl₃) δ: 4.44 (2H, s), 7.60-7.90 (3H, m), 8.11 (1H, s),

FD MS (m/z): 263 (M⁺), 265 (M⁺)

b) 444 mg of selenium dioxide was dissolved in 10 ml of dry methanolwith heating, followed by the addition of 1.056 g of2-(2-bromo-1-oxoethyl)-5-benzofurancarbonitrile obtained in the abovestep a). The thus prepared mixture was refluxed under heating for 12hours. After cooling, insoluble materials were removed by filtration,and the resulting filtrate was concentrated to dryness. Thereafter, theresulting residue was purified by subjecting it to silica gel columnchromatography using a toluene/ethyl acetate mixture as an eluant,thereby obtaining 129 mg of methyl2-(5-cyano-2-benzofuranyl)-2-oxoacetate in the form of colorless needlecrystals.

mp: 196°-199° C.

IR (KBr): 1740, 1674, 1614, 1552 cm⁻¹

¹ H-NMR (CDCl₃) δ:4.03 (3H, s), 7.66-7.96 (2H, m), 8.17 (2H, s)

FD MS (m/z): 321 (M⁺ +92), 229 (M⁺)

c) 3.1 g of methyl 2-(5-cyano-2-benzofuranyl)-2-oxoacetate obtained inthe above step b) and 6.2 g of(4-methoxyphenyl)methyltriphenylphosphonium chloride were dissolved in amixture solvent of 100 ml of tetrahydrofuran and 100 ml of methanol.With stirring at room temperature, 2.19 g of1,8-diazabicyclo[5.4.0]-7-undecene was added to the above solution, andthe stirring was continued for 1 hour. To this were further added 1.3 gof (4-methoxyphenyl)methyltriphenylphosphonium chloride and 0.65 g of1,8-diazabicyclo[5.4.0]-7-undecene. After stirring for 1 hour andsubsequently removing the solvent by distillation, the resulting residuewas purified by subjecting it to silica gel column chromatography usingchloroform as an eluant, thereby obtaining a viscous and oily olefiniccompound as a mixture of E and Z forms.

¹ H-NMR (CDCl₃) δ:3.78 (1.5H, s), 3.84 (3H, s), 3.87 (1.5H, s), 6.60(9H, m)

The olefinic compound obtained above was dissolved in a solvent mixtureof 100 ml of tetrahydrofuran and 100 ml of methanol, followed by theaddition of 1.1 g of palladium.1H₂ O.bariumsulfate and by subsequentcatalytic hydrogenation under normal pressure for 3 hours. Afterremoving the catalyst by filtration and distilling off the solvent, theresulting residue was purified by subjecting it to silica gel columnchromatography to obtain 4.2 g of viscous and oily methyl2-(5-cyano-2-benzofuranyl)-3-(4-methoxyphenyl)propionate.

¹ H-NMR (CDCl₃) δ:3.20 (1H, dd, J=14.4 and 7.8Hz), 3.41 (1H, dd, J=14.4and 7.4Hz), 3.69 (3H, s), 3.75 (3H, s), 4.10 (1H, dd, J=7.8 and 7.4Hz),6.60 (1H, s), 6.76 (2H, d, J=8.8Hz), 7.05 (2H, d, J=8.8Hz), 7.53 (2H),7.82 (1H, s)

d) 4.2 g of methyl2-(5-cyano-2-benzofuranyl)-3-(4-methoxyphenyl)propionate obtained in theabove step c) was dissolved in 150 ml of dichloromethane, and thesolution was cooled down to -50° C. With stirring, to this was addeddropwise 30 ml of a dichloromethane solution containing 9.97 g of borontribromide. The temperature of the resulting reaction solution wasgradually increased to 15° C. After 30 minutes of stirring at thistemperature, the reaction solution was diluted with chloroform, washedwith dilute hydrochloric acid, and then dried to distill off thesolvent. The resulting residue was subjected to silica gel columnchromatography using a chloroform/ethanol mixture as an eluant, and thesolvent in pooled fractions of interest was concentrated to precipitatecrystals. The crystals thus precipitated were washed with benzene, andcollected by filtration, thereby obtaining 3.1 g of methyl2-(5-cyano-2-benzofuranyl)-3-(4-hydroxyphenyl)propionate in the form ofcolorless crystals.

mp: 110°-111° C.

IR (KBr): 2228, 1722, 1594, 1518, 1272 cm⁻¹

¹ H-NMR (CDCl₃) δ: 3.18 (1H, dd, J=14.4 and 7.8Hz), 3.36 (1H, dd, J=14.4and 7.4Hz), 3.69 (3H, s), 4.09 (1H, dd, J=7.8 and 7.4Hz), 6.60 (1H, s),6.69 (2H, d, J=8.4Hz), 7.00 (2H, d, J=8.4Hz), 7.53 (2H, s), 7.83 (1H, s)

e) In 150 ml of dry tetrahydrofuran were dissolved 3.0 g of methyl2-(5-cyano-2-benzofuranyl)-3-(4-hydroxyphenyl)propionate obtained in theabove step d), 1.92 g of (3R)-1-tert-butoxycarbonyl-3-hydroxypyrrolidineand 2.69 g of triphenylphosphine. With stirring at room temperature,1.79 g of diethyl azodicarboxylate was added to the thus preparedsolution, and the stirring was continued for 1 hour. After distillingoff the solvent, the resulting residue was subjected to silica gelcolumn chromatography using a toluene/ethyl acetate mixture as aneluant, thereby obtaining a mixture consisting of the starting materialmethyl 2-(5-cyano-2-benzofuranyl)-3-(4-hydroxyphenyl)propionate and thetitle compound.

The mixture thus obtained was dissolved in 100 ml of tetrahydrofuran. Tothis were added 0.95 g of(3R)-1-tert-butoxycarbonyl-3-hydroxypyrrolidine, 1.35 g oftriphenylphosphine and 0.85 g of diethyl azodicarboxylate in that order.The resulting mixture was stirred at room temperature for 16 hours.Thereafter, the resulting reaction solution was treated and purified inthe same manner as described above to obtain 2.02 g of the titlecompound in a viscous and oily form.

¹ H-NMR (CDCl₃) δ:1.46 (9H, s), 1.88-2.24 (2H, m), 3.10-3.60 (6H, m),3.69 (3H, s), 4.10 (1H, t), 4.81 (1H, br), 6.61 (1H, s), 6.73 (2H, d,J=8.3Hz), 7.04 (2H, d, J=8.3Hz), 7.54 (2H, s), 7.83 (1H, s)

FD MS (m/z): 321 (M⁺)

REFERENCE EXAMPLE 66 Preparation of ethyl3-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-4-5-cyanobenzo[b]thien-2-yl)-butyrate

a) 14.2 g of ethyl 2-ethoxycarbonyl-2-(4-methoxyphenyl)acetate wasdissolved in 150 ml of tetrahydrofuran. 2.6 g of sodium hydride (oiltype, 60%) was added to the above solution while stirring under icecooling, and the stirring was continued for 20 minutes. To this wasfurther added 17.2 g of 5-bromo-2-bromomethylbenzo[b]thiophene. Afterstirring at room temperature for 18 hours, the resulting reactionsolution was mixed with ammonium chloride aqueous solution, and thenextracted with ethyl acetate. After drying to distill off the solvent,the resulting residue was subjected to silica gel column chromatographyusing chloroform as an elution solvent, thereby obtaining 24.2 g ofethyl3-(5-bromobenzo[b]thien-2-yl)-2-ethoxycarbonyl-2-(4-methoxyphenyl)propionate.

¹ H-NMR (CDCl₃) δ:1.2 (6H, t), 3.78 (3H, s), 3.85 (2H, s), 6.75-7.0 (3H,m), 7.2-7.8 (5H, m)

b) A solution of 7.3 g of potassium hydroxide dissolved in 20 ml ofwater was added to 200 ml of an ethanol solution containing 24.2 g of3-(5-bromobenzo[b]thien-2-yl)-2-ethoxycarbonyl-2-(4-methoxyphenyl)propionateobtained in the above step a), and the thus prepared mixture was stirredfor 4 days. The resulting reaction solution was poured into cooleddilute hydrochloric acid to collect precipitated crystals by filtration.The thus collected crystals were dissolved in ethyl acetate, and thendried. After distilling off the solvent, the resulting residue wasdissolved in 200 ml of ethanol, mixed With 3 ml of concentrated sulfuricacid, and then refluxed under heating for 2 hours. After cooling, theresulting reaction solution was concentrated, mixed with chloroform,washed with water, and then dried. After distilling off the solvent, theresulting residue was purified by subjecting it to silica gel columnchromatography using chloroform as an eluant, thereby obtaining 20 g ofethyl 3-(5-bromobenzo[b]thien-2-yl)-2-(4-methoxyphenyl)propionate.

¹ H-NMR (CDCl₃) δ: 1.17 (3H, t), 3.2 (1H, dd), 3.55 (1H, dd), 3.77 (3H,s), 3.81 (1H, dd), 4.10 (2H, q), 6.82 (2H, d), 7.2-7.8 (6H, m)

c) 20 g of ethyl3-(5-bromobenzo[b]thien-2-yl)-2-(4-methoxyphenyl)propionate obtained inthe above step b) was dissolved in 200 ml of tetrahydrofuran, followedby the addition of 12 g of sodium borohydride. To the mixture was addeddropwise 80 ml of methanol under ice cooling. After stirring for 3hours, the resulting reaction solution was adjusted to pH 6 withconcentrated hydrochloric acid, and then extracted with ethyl acetate.The resulting organic layer was dried to distill off the solvent, andthe thus obtained residue was subjected to silica gel columnchromatography using a chloroform/methanol mixture as an elution solventto obtain 16 g of3-(5-bromobenzo[b]thien-2-yl)-2-(4-methoxyphenyl)-1-propanol.

¹ H-NMR (CDCl₃) δ:2.9-3.4 (3H, m), 3.73 (3H, s), 3.62-3.90 (2H, br),6.70-7.80 (8H, m)

d) 16 g of 3-(5-bromobenzo[b]thien-2-yl)-2-(4-methoxyphenyl)-1-propanolobtained in the above step c) was dissolved in 40 ml of dichloromethane.To the mixture were added 6.3 ml of triethylamine and 4 ml ofmethanesulfonyl chloride while stirring under ice cooling. Afterstirring at the same temperature for 2 hours, the resulting reactionsolution was mixed with dichloromethane, washed with water and thendried. After distilling off the solvent, the resulting residue waspurified by subjecting it to silica gel column chromatography usingchloroform as an elution solvent, thereby obtaining 18.5 g of3-(5-bromobenzo[b]-thien-2-yl)-2-(4-methoxyphenyl)propylmethanesulfonate.

¹ H-NMR (CDCl₃) δ: 3.78 (3H, s), 3.9-4.5 (3H, m), 3.70 (3H, s), 4.3 (2H,m), 6.70-7.80 (8H, m)

e) 1.2 g of sodium cyanide was dissolved in 30 ml of dimethyl sulfoxideat a temperature of 90° C. To this was gradually added 18.5 g of3-(5-bromobenzo[b]thien-2-yl)-2-(4-methoxyphenyl)propylmethanesulfonate, followed by stirring at 80° C. for 1 hour. Theresulting reaction solution was mixed with an ethyl acetate/toluenemixture, washed with water, and then dried to distill off the solvent.Crystals thus precipitated were washed with ethanol and dried to obtain5 g of 3-(5-bromobenzo[b]thien-2-yl)-2-(4-methoxyphenyl)butyronitrile.The same compound was also obtained with a yield of 2 g, byconcentrating the ethanol solution resulting from the washing ofcrystals and subjecting the concentrate to silica gel columnchromatography using chloroform as an elution solvent.

¹ H-NMR (CDCl₃) δ:2.5-2.7 (2H, br), 3.2-3.4 (3H, br), 3.76 (3H, s),6.70-7.80 (8H, m)

MS m/z: 386, 388

f) 7 g of 3-(5-bromobenzo[b]thien-2-yl)-2-(4-methoxyphenyl)butyronitrileobtained in the above step e) was suspended in 80 ml of ethanol,followed by the addition of 5 ml of concentrated sulfuric acid and a fewdrops of water. The thus prepared mixture was refluxed under heating for7 days. After distilling off the solvent, the thus obtained reactionsolution was mixed with chloroform and water, and the resulting organiclayer was dried to distill off the solvent. Thereafter, the resultingresidue was subjected to silica gel column chromatography usingchloroform as an elution solvent to obtain 6.3g of ethyl4-(5-bromobenzo[b]thien-2-yl)-3-(4-methoxyphenyl)butyrate.

¹ H-NMR (CDCl₃) δ: 1.12 (3H, t), 2.65 (2H, dd), 3.10-3.80 (3H, m), 3.76(3H, s), 4.01 (2H, q), 6.70-6.95 (3H, m), 7.10 (2H, d), 7.20-7.40 (1H),7.55 (1H, d), 7.72 (1H, d)

FAB MS (m/z): 433, 435

g) 6.0 g of ethyl4-(5-bromobenzo[b]thien-2-yl)-3-(4-methoxyphenyl)butyrate obtained inthe above step f) was dissolved in 50 ml of N-methyl-2-pyrrolidone,followed by the addition of 1.6 g of cuprous cyanide and a catalyticallyeffective amount of copper sulfate. The mixture thus prepared wasstirred at a temperature of 190° to 200° C. in a stream of argon. Aftercooling, the resulting reaction solution was mixed with ethyl acetateand toluene and then washed with water, followed by drying to distilloff the solvent. Thereafter, the resulting residue was purified bysubjecting it to silica gel column chromatography using achloroform/acetone mixture as an elution solvent, thereby obtaining 4.5g of ethyl 4-(5-cyanobenzo[b]thien-2-yl)-3-(4-methoxyphenyl)butyrate.

¹ H-NMR (CDCl₃) δ: 1.18 (3H, t), 2.70 (2H, dd), 3.16-3.70 (3H, m), 3.78(3H, s), 4.02 (2H, q), 6.85 (2H, d), 6.98 (1H, s), 7.18 (2H, d), 7.5(1H, dd), 7.8 (1H, d), 7.96 (1H, d)

h) 4.5 g of ethyl4-(5-cyanobenzo[b]thien-2-yl)-3-(4-methoxyphenyl)butyrate obtained inthe above step g) was dissolved in 20 ml of dichloromethane. 3.4 ml ofboron tribromide was added to the above solution which has been cooleddown to -70° C. The thus prepared mixture was warmed up to roomtemperature, and stirred for 1 hour. Chipped ice was added to theresulting reaction solution to collect a dichloromethane layer which wassubsequently dried to distill off the solvent. The resulting residue wasdissolved in 50 ml of tetrahydrofuran. With stirring under ice coolingin a stream of argon, 1.9 g of(3R)-1-tert-butoxycarbonyl-3-hydroxypyrrolidine, 3.2 g oftriphenylphosphine and 2.3 g of diethyl azodicarboxylate were added tothe resulting mixture. The thus prepared mixture was stirred at roomtemperature for 18 hours. After distilling off the solvent, theresulting residue was purified by subjecting it to silica gel columnchromatography using a n-hexane/ethyl acetate mixture as an elutionsolvent. In this way, 4 g of the title compound was obtained.

¹ H-NMR (CDCl₃) δ:1.48 (9H, s), 1.95-2.20 (2H, m), 2.65 (2H, dd),3.15-3.70 (7H, m), 4.78-5.00 (1H, m), 6.80 (2H, d), 6.98 (1H, s), 7.17(2H, d), 7.5 (1H, dd), 7.82 (1H, d), 7.98 (1H, d)

REFERENCE EXAMPLE 67 Preparation of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)thio]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)-2-yl)-2-ethoxycarbonylpropionate

a) 20.2 g of ethyl 4-mercaptophenylacetate was dissolved in 450 ml oftetrahydrofuran. With ice cooling and with stirring, 21.0 g of(3R)-1-tert-butoxycarbonyl-3-hydroxypyrrolidine, 29.4 g oftriphenylphosphine and 19.5 g of diethyl azodicarboxylate were added tothe above solution. The thus prepared mixture was stirred at roomtemperature for 18 hours. After distilling off the solvent, theresulting residue was purified by subjecting it to silica gel columnchromatography using a n-hexane/ethyl acetate mixture as an elutionsolvent, thereby obtaining 7.0 g of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)thio]phenyl]acetate.

¹ H-NMR (CDCl₃) δ:1.25 (3H, t, J=7.2Hz), 1.45 (9H, s), 1.7-2.4 (2H, m),3.2-4.4 (5H, m), 3.58 (2H, s), 4.15 (2H, q, J=7.2Hz), 7.0-7.6 (4H, m)

b) 4.0 g of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)thio]phenyl]acetateobtained in the above step a) was dissolved in 21 ml ofN,N-dimethylformamide, followed by the addition of 4.02 ml of diethylcarbonate. With stirring at a temperature of 130° C., 530 mg (60%) ofsodium hydride was added to the thus prepared solution, and theresulting mixture was stirred for 10 minutes, followed by further adding106 mg of sodium hydride and by additional stirring for 10 minutes. Theresulting reaction solution was poured into ice water, neutralized withdilute hydrochloric acid, extracted with ethyl acetate, and then dried.After distilling off the solvent, the resulting residue was purified bysubjecting it to silica gel column chromatography using a n-hexane/ethylacetate mixture as an elution solvent, thereby obtaining 1.74 g of ethyl2-[4-[((3S)-1-tert-butoxy-carbonyl-3-pyrrolidinyl)thio]phenyl]-2-ethoxycarbonylacetatein an oily form.

¹ H-NMR (CDCl₃) δ:1.27 (6H, t, J=7.2Hz), 1.46 (9H, s), 1.4-2.4 (2H, m),3.0 -4.0 (5H, m), 4.22 (4H, q, J=7.2Hz), 4.58 (1H, s), 7.2-7.5 (4H, m)

c) 1.7 g of ethyl2-[4-[((3S)-1-tert-butoxy-carbonyl-3-pyrrolidinyl)thio]phenyl]-2-ethoxycarbonylacetate obtained in the above step b) was dissolved in a solvent mixtureof 20 ml of tetrahydrofuran and 1 ml of N,N-dimethylformamide, followedby the addition of 155 mg of sodium hydride (60%) and by subsequentstirring for 20 minutes. 980 mg of2-bromomethylbenzo[b]thiophene-5-carbonitrile was added to the abovereaction solution, and the mixture was stirred for 24 hours. Theresulting reaction solution was poured into ice water, extracted withethyl acetate, and then dried. After distilling off the solvent, theresulting residue was purified by subjecting it to silica gel columnchromatography using a n-hexane/ethyl acetate mixture as an elutionsolvent, thereby obtaining 2.05 g of the title compound in an oily form.

¹ H-NMR (CDCl₃) δ:1.23 (6H, t, J=7.2Hz), 1.46 (9H, s), 1.50-2.50 (2H,m), 3.2-4.4 (5H, m), 3.89 (2H, s), 4.25 (4H, q, J=7.2Hz), 7.28 (4H, s),7.44 (1H, dd, J=8.4 and 1.5Hz), 7.78 (1H, d, J=8.4Hz), 7.91 (1H, dd)

REFERENCE EXAMPLE 68 Preparation of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)-2-propionate

In a solvent mixture of 50 ml of tetrahydrofuran and 50 ml of ethanolwere dissolved 3.0 g of(5-cyanobenzo[b]thien-2-yl)methyltriphenylphosphoniumchloride and 2.55 gof ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-2-oxoacetate.With stirring at room temperature, 1.07 g of1,8-diazabicyclo[5.4.0]-7-undecene was added to the thus preparedsolution, and the resulting mixture was stirred for 1 hour at roomtemperature. After distilling off the solvent, the resulting residue waspurified by subjecting it to silica gel column chromatography using atoluene/ethyl acetate mixture as an eluant, thereby obtaining ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)acrylateas a mixture of E and Z forms. The thus obtained compound was dissolvedin a solvent mixture of 50 ml of tetrahydrofuran and 50 ml of ethanol,and the resulting solution was mixed with 5.0 g of 10% palladium carboncatalyst (50% wet type), and subjected to catalytic hydrogenation undernormal pressure. After removing the catalyst by filtration anddistilling off the solvent, the resulting residue was purified bysubjecting it to silica gel column chromatography using a toluene/ethylacetate mixture as an eluant, thereby obtaining 2.2 g of the titlecompound in a viscous and oily form.

¹ H-NMR (CDCl₃) δ: 1.17 (3H, t, J=7.0Hz), 1.47 (9H, s), 1.90-2.20 (2H,m), 3.10-3.95 (7H, m), 4.10 (2H, q, J=7.0Hz), 4.84 (1H, br), 6.81 (2H,d, J=9.0Hz), 7.20 (1H, s), 7.25 (2H, d, J=9.0Hz), 7.44 (1H, dd, J=9.0and 1.6Hz), 7.81 (1H, dd, J=9.0 and 1.6Hz), 7.94 (1H, s)

The following compounds of Reference Examples 69 to 75 were prepared inaccordance with the procedure described in Reference Example 68.

REFERENCE EXAMPLE 69 ethyl2-[4-[2-(tert-butoxycarbonylamino)-1-tert-butoxycarbonylaminomethyl)ethoxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)propionate

viscous oil

¹ H-NMR (CDCl₃) δ: 1.25 (3H, t, J=7.0Hz), 1.45 (18H, s), 2.90-4.50 (8H,m), 6.80-7.35 (5H), 7.45 (1H, dd, J=8.3 and 1.3Hz), 7.80 (1H, d,J=8.3Hz), 7.93 (1H)

REFERENCE EXAMPLE 70 ethyl2-[4-[((2S)-1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)propionate

¹ H-NMR (CDCl₃) δ:1.17 (3H, t), 1.47 (9H, s), 2.00 (4H, br), 3.40 (2H,br), 3.60-4.30 (6H), 6.90 (2H, d, J=10Hz), 7.25 (2H, d, J=10Hz),7.00-8.00 (4H, m)

REFERENCE EXAMPLE 71 ethyl2-[4-[(1-tert-butoxycarbonyl-4-piperidinyl)oxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)propionate

solid

¹ H-NMR (CDCl₃) δ: 1.10 (3H, t, J=6.0Hz), 1.50 (9H, s), 1.70-2.00 (4H,m), 3.20-4.00 (4H, m), 4.15 (2H, q), 4.30-4.60 (1H, br), 6.80-8.10 (8H)

REFERENCE EXAMPLE 72 ethyl2-[4-(2-tert-butoxycarbonylaminoethoxy)phenyl]-3-(5-cyanobenzo[b]thien-2-yl)propionate

viscous oil

¹ H-NMR (CDCl₃) δ:1.16 (3H, t, J=7.0Hz), 1.45 (9H, s), 3.05-4.40 (9H),5.12 (1H, br), 6.84 (2H, d, J=8.3Hz), 7.01 (1H, s), 7.25 (2H, d,J=8.3Hz), 7.41 (1H, dd, J=8.3 and 1.2Hz), 7.77 (1H, d, J=8.3Hz), 7.89(1H, s)

REFERENCE EXAMPLE 73 ethyl2-[4-[((2S)-1-tert-butoxycarbonyl-5-oxo-2-pyrrolidinyl)methoxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)propionate

viscous oil

¹ H-NMR (CDCl₃) δ:1.17 (3H, t), 1.42 (9H,s), 1,80-2.25 (2H, m),2.30-2.60 (2H, m), 3.20 (1H, dd), 3.37 (1H, dd), 3.50-3.82 (1H, dd),3.82-4.50 (4H, m), 4.80-5.10 (1H, m), 6.75-8.10 (8H, m)

REFERENCE EXAMPLE 74 ethyl 2-[4-[((2R,4S)-1-tert-butoxycarbonyl-2-methyl-4-pyrrolidinyl)oxy]phenyl]-3-(6-cyanobenzo[b]thien-2-yl)propionate

¹ H-NMR (CDCl₃) δ:1.15-1.50 (6H, m), 1.50 (9H, s), 1.80-2.60 (2H, m),3.00-4.50 (8H, m), 4.80-5.10 (1H, m), 6.80-8.20 (8H, m)

REFERENCE EXAMPLE 75 ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyanobenzo[b]thien-2-yl)propionate

¹ H-NMR (CDCl₃) β:1.17 (3H, t), 1.46 (9H, s), 2.10 (2H, m), 3.60 (6H,m), 3.83 (1H, m), 4.10 (2H, q), 4.85 (1H, br), 6.86 (2H, d), 7.04 (1H,s), 7.25 (2H), 7.55 (1H, dd), 7.65 (1H, d), 8.04 (1H)

REFERENCE EXAMPLE 76 Preparation of ethyl(+)-2-[4-[((2S)-1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)propionateand ethyl(-)-2-[4-[((2S)-3-pyrrolidinyl)methoxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)propionate

5.0 g of ethyl2-[4-[((2S)-1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)propionatewas separated into (+) and (-) forms using a column for optical isomerseparation, thereby obtaining 2.5 g (+) and 1.7 g (-) of the titlecompounds.

(-) form:

mp: 102°-104° C.

[α]_(D) ²⁴ =-142.00 (c=1,000, EtOH)

¹ H-NMR (CDCl₃) δ:1.13-1.22 (3H, m), 1.47 (9H, s), 1.80-2.10 (4H, m),3,25-3.50 (4H, m), 3.64-3.75 (1H, m), 3.70-3.90 (1H, br), 3.90 (1H, br),4.05-4.20 (4H, m), 6.88 (2H, d, J =8.3Hz), 7.02 (1H, s), 7.23 (2H, d,J=8.3Hz), 7.45 (1H, d, J=8.3Hz), 7.80 (1H, d, J=8.3Hz), 7.94 (1H, s)

HPLC: Column; an amylose-based column for use in the separation ofoptical isomers (CHIRALPAK AD, 20 φ×250 mm, Daicel Chemical Industries,Ltd.)

Solvent; n-hexane:iso-propanol=70:30

Flow rate; 4 ml/min

Retention time; 20 to 23 minutes

(+) form:

mp: 111°-112° C.

[α]_(D) ²⁴ =+55.19 (c=1.000, EtOH)

¹ H-NMR (CDCl₃) δ:1.13-1.22 (3H, m), 1.47 (9H, s), 1.80-2.10 (4H, m),3.25-3.50 (4H, m), 3.64-3.75 (1H, m), 3.70-3.90 (1H, br), 3.90 (1H, br),4.05-4.20 (4H, m), 6.88 (2H, d, J =8.3Hz), 7.02 (1H, s), 7.23 (2H, d,J=8.3Hz), 7.45 (1H, d, J=8.3Hz), 7.80 (1H, d, J=8.3Hz), 7.94 (1H, s)

HPLC: Column; an amylose-based column for use in the separation ofoptical isomers (CHIRALPAK AD, 20 φ×250 mm, Daicel Chemical Industries,Ltd.)

Solvent; n-hexane:iso-propanol=70:30

Flow rate; 4 ml/min

Retention time; 23 to 27 minutes

REFERENCE EXAMPLE 77 Preparation of ethyl (-)-2-[4-[(1-tert-butoxycarbonyl-4-piperidinyl)oxy]phenyl]-3-(7-cyano-2-naphthyl)propionateand ethyl(+)-2-[4-[(1-tert-butoxycarbonyl-4-piperidinyl)oxy]phenyl]-3-(7-cyano-2-naphthyl)propionate

These compounds were prepared in the same manner as described inReference Example 76.

(-) form:

[α]_(D) ²⁴ =-100.78° (c=1.024, CHCl₃)

¹ H-NMR (CDCl₃) δ:1.11 (3H, t, J=6.9Hz), 1.47 (9H, s), 1.70-1.80 (2H,m), 1.85-1.95 (2H, m), 3.15-3.20 (1S, m), 3.30-3.40 (2H, m), 3.50-3.60(1H, m), 3.65-3.75 (2H, m), 3.85-3.90 (1H, br), 4.0-4.1 (2H, m),4.40-4.45 (1H, m), 6.85 (2H, d, J=8.3Hz), 7.23 (2H, d), 7.40-7.45 m),7.53-7.58 (1H, m), 7.62 (1H, s), 7.77 (2H, d), 7.85 (1H, d, J=8.3Hz),8.12 (1H, s)

HPLC: Column; an amylose-based column for use in the separation ofoptical isomers (CHIRALPAK AD, 4.6 φ×250 mm, Daicel Chemical Industries,Ltd.)

Solvent; n-hexane:iso-propanol=90:10

Flow rate; 1 ml/min

Retention time; 26.9 minutes

(+) form:

[α]_(D) ²⁴ =+95.84° (c=1.010, CHCl₃)

¹ H-NMR (CDCl₃) δ:1.11 (3H, t, J=7.3Hz), 1.65-1.70 (2H, m), 1.85-2.00(2H, m), 3.15-3.20 (1H, m), 3.30-3.35 (2H, m), 3.50-3.60 (1H, m),3.65-3.75 (2H, m), 3.85-3.90 (1H, br),4.0-4.1 (2H, m), 4.40-4.45 (1H,m), 6.85 (2H, d, J=8.8Hz), 7.23 (2H, d, J=8.3Hz), 7.40-7.45 (m, 1H,Ar-H), 7.52-7.57 (1H, m), 7.62 (1H, s), 7.77 (1H, d, J=8.3Hz), 7.85 (1H,d, J=8.3Hz), 8.11(1H, s)

HPLC: Column; an amylose-based column for use in the separation ofoptical isomers (CHIRALPAK AD, 4.6 φ×250 mm, Daicel Chemical Industries,Ltd.)

Solvent; n-hexane:iso-propanol=90:10

Flow rate; 1 ml/min

Retention time; 31 minutes

REFERENCE EXAMPLE 78 Preparation of ethyl(+)-2-[4-[((2S)-1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)propionate

54.0 g of2-[4-[((2S)-1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)propionatewas dissolved in 400 ml of dry ethanol during heating, and 800 ml of dryn-hexane was added to the resulting solution. To the thus preparedmixture were added 100 mg of sodium hydride and seed crystals of ethyl(+)-2-[4-[((2S)-1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)propionate.After stirring at room temperature for 4 hours, the thus stirred mixturewas further mixed with 100 mg of sodium hydride, and the stirring wascontinued for additional 18 hours at room temperature, followed bycollecting precipitated crystals by filtration. The thus collectedcrystals were recrystallized from 22 volumes (w/v) of anethanol/n-hexane mixture (30:70, w/v). By repeating therecrystallization step three times, 37.0 g of the title compound wasobtained with a diastereoisomer purity of 99.5% or more.

REFERENCE EXAMPLE 79 Preparation of ethyl(+)-2-[4-[((3S)-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-cyano-2-naphthyl)propionate

This compound was obtained in similar manner to the procedure ofReference Example 78.

REFERENCE EXAMPLE 80 Preparation of ethyl2-[4-[((2R)-1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)-2-ethoxycarbonylpropionate

4.1 g of ethyl2-[4-[((2R)-1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxy]phenyl]-2-ethoxycarbonylacetatewas dissolved in 100 ml of tetrahydrofuran. At room temperature, thethus prepared solution was mixed with 0.38 g of 60% sodium hydride andstirred for 30 minutes. With stirring at room temperature, to theresulting reaction solution was added dropwise 10 ml of atetrahydrofuran solution containing 2.1 g of2-bromomethylbenzo[b]-thiophene-5-carbonitrile. After concentrating thereaction solution to dryness, the resulting residue was purified bysubjecting it to silica gel column chromatography using atoluene/chloroform mixture as an elution solvent, thereby obtaining 4.34g of the title compound in an oily form.

¹ H-NMR (CDCl₃) δ:1.21 (6H), 1.46 (9H, s), 2.0 (4H, br), 3.40 (2H, br),3.88 (3H), 4.22 (6H), 6.90 (3H), 7.20 (2H, d), 7.50 (1H), 7.78 (1H, d),7.93 (1H, d)

The following compounds of Reference Examples 81 and 82 were prepared inaccordance with the procedure described in Reference Example 80.

REFERENCE EXAMPLE 81 ethyl3-(5-cyanobenzo[b]thien-2-yl)-2-ethoxycarbonyl-2-[4-[(2-imidazolin-2-yl)methoxy]phenyl]propionate

viscous oil

¹ H-NMR (CDCl₃) δ:1.22 (6H, t), 3.63 (4H, s), 3.89 (2H, s), 4.24 (4H),4.69 (2H, s), 6.86 (3H), 7.27 (2H), 7.42 (1H), 7.76 (1H), 7.88 (1H)

REFERENCE EXAMPLE 82 ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-benzothiazolyl)-2-ethoxycarbonylpropionate

viscous oil

¹ H-NMR (CDCl₃) 1.22 (6H, t), 1.46 (9H, s), 2.09 (2H, br), 3.56 (4H,br), 4.13 (2H), 4.28 (4H, q), 4.85 (1H, br), 6.82 (2H, d), 7.26 (2H, d),7.63 (1H, dd), 7.95 (1H, d), 8.25 (1H, d)

REFERENCE EXAMPLE 83 Preparation of ethyl3-(5-cyanobenzo[b]thien-2-yl)-2-[4-[2-(ethoxycarbonylimino)hexahydropyrimidine-5-yl]oxy]phenyl]propionate

1.0 g of ethyl 2-[4-[2-(tert-butoxycarbonylamino)-1-(tert-butoxycarbonylaminomethyl)ethoxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)propionatewas dissolved in 2 ml of anisole. 10 ml of trifluoroacetic acid wasadded to the above solution while stirring under ice cooling, and thethus prepared mixture was stirred-at room temperature for 1 hour. Theresulting reaction solution was concentrated under a reduced pressure,and the thus obtained residue was dissolved in water and washed withn-hexane. The resulting water layer was adjusted to pH 9-10 withconcentrated aqueous ammonia and then extracted with chloroform. Theresulting organic layer was concentrated to dryness, and the thusobtained residue was dissolved in 20 ml of dry ethanol. To the thusprepared solution was added 300 mg of ethylN-(ethoxy(methylthio)methylene) carbamate which has been synthesized inaccordance with the procedure disclosed in Journal of the ChemicalSociety, Parkin I, 1973, pp. 2644-2646. Thereafter, the thus preparedmixture was stirred for 20 hours, and the resulting precipitate wascollected to obtain 560 mg of the title compound.

mp: 179°-182° C.

IR (KBr): 2230, 1725, 1638, 1512, 1337 cm⁻¹

¹ H-NMR (CDCl₃) δ:1.13 (3H, t, J=7.0Hz), 1.17 (3H, t, J=7.0Hz),3.10-4.30 (11H, m), 4.50-4.80 (1H, m), 6.89 (2H, d, J=8.75Hz), 7.03 (1H,s), 7.26 (2H, d, J=8.75Hz), 7.46 (1H, dd, J=8.31 and 1.75Hz), 7.82 (1H,d, J=8.31Hz), 7.95 (1H, d), 8.70-9.50 (2H, br)

REFERENCE EXAMPLE 84 Preparation of ethyl3-(5-cyanobenzo[b]thien-2-yl)-2-[4-[[2-(imino)hexahydropropyrimidin-5-yl]oxy]phenyl]propionatehydrochloride

a) 2.9 g of potassium thiocyanate was dissolved in 150 ml of dryacetone. With stirring under ice cooling, to the above solution wasadded dropwise 6.8 g of p-nitrobenzyl chloroformate which had beendissolved in 20 ml of acetone. The thus prepared mixture was stirred for2 hours with ice cooling, and the resulting mixture was mixed with 1.15g of methanol, and stirred at room temperature for 20 hours. Thereafter,crystals thus precipitated were collected by filtration and washed withchloroform to obtain 2.88 g of p-nitrobenzyl methoxy(thiocarbamoyl)carbamate in the form of powder.

¹ H-NMR (CDCl₃) δ:4.03 (3H, s), 5.33 (2H, s), 7.70 (2H, d, J=9.0Hz),8.80 (2H, d, J=9.0Hz)

b) 3.5 g of p-nitrobenzyl methoxy(thiocarbamoyl) carbamate obtained inthe above step a) and 1.79 g of anhydrous potassium carbonate weredissolved in a mixture solution of 40 ml of water and 40 ml of dioxane.1.72 g of dimethyl sulfate was gradually added dropwise to the thusprepared solution, and the resulting mixture was stirred at roomtemperature for 30 minutes. To the resulting reaction solution was addedagain 300 mg of anhydrous potassium carbonate, followed by the dropwiseaddition of 300 mg of dimethyl sulfate. The reaction solution thusobtained was diluted with ethyl acetate, washed with water and saturatedsodium chloride aqueous solution in that order, and then concentrated.Thereafter, crystals thus precipitated were collected by filtration, andwashed thoroughly with n-pentane to obtain 3.23 g of p-nitrobenzylN-(methoxy(methylthio)methylene) carbamate.

¹ H-NMR (CDCl₃) δ: 2.40 (3H, s), 4.00 (3H, s), 5.28 (2H, s), 7.56 (2H,d, J=9.0Hz), 8.22 (2H, d, J=9.0Hz)

c) In 10 ml portion of anisole was dissolved 2.0 g of ethyl2-[4-[2-(tert-butoxycarbonylamino)-1-(tertbutoxycarbonylaminomethyl)ethoxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)propionate.30 ml of trifluoroacetic acid was added to the above solution duringstirring under ice cooling, and the mixture was stirred at roomtemperature for 2 hours. The resulting reaction solution wasconcentrated under a reduced pressure, and the thus obtained residue wasdissolved in water, and washed with n-hexane. The resulting water layerwas adjusted to pH 10 with concentrated aqueous ammonia and thenextracted with chloroform. The resulting organic layer was concentratedto dryness, and the thus obtained residue was dissolved in 50 ml oftetrahydrofuran. To the thus prepared solution was added 921 mg ofp-nitrobenzyl N-(methoxy(methyl-thio)methylene) carbamate obtained inthe above step b), followed by stirring for 18 hours. After distillingoff the solvent, the resulting residue was purified by subjecting it tosilica gel column chromatography using a chloroform/ethanol mixture asan elution solvent, thereby obtaining 1.5 g of ethyl3-(5-cyanobenzo[b]thien-2-yl)-2-[4-[[2-(p-nitrobenzyloxycarbonylimino)hexahydropyrimidin-5-yl]oxy]phenyl]propionatein an viscous and oily form.

¹ H-NMR (CDCl₃) δ:1.17 (3H, t, J=7.0Hz), 3.00-4.30 (1H, m), 4.40-4.70(1H, m), 5.08 (2H, s), 6.81 (2H, d, J=8.3Hz), 7.03 (1H, s), 7.10-7.56(5H, m), 7.81 (1H, d, J=9.3Hz), 7.94 (1H, s), 8.10 (2H, d, J=8.75Hz),8.70-9.40 (2H, br)

d) In 100 ml portion of ethanol was dissolved 1.5 g of ethyl3-(5-cyanobenzo[b]thien-2-yl)-2-[4-[[2-(p-nitrobenzyloxycarbonylimino)hexahydropyrimidin-5-yl]oxy]phenyl]propionateobtained in the above step c). To the thus prepared solution were added0.5 g of ammonium chloride and 0.5 g of 10% palladium carbon catalyst(50% wet type). The resulting mixture was subjected to 2 hours ofcatalytic hydrogenation under normal pressure. After removing thecatalyst by filtration and distilling off the solvent, the resultingresidue was purified by subjecting it to silica gel columnchromatography using a chloroform/ethanol mixture as an elution solvent,thereby obtaining 1.0 g of the title compound.

¹ H-NMR (CDCl₃) δ: 1.21 (3H, t, J=7.0Hz), 3.00-3.90 (7H, m), 4.17 (2H,q, J=7.0Hz), 4.50-4.80 (1H, br), 6 87 (2H, d, J=8.75Hz), 7.01 (1H, s),7.06-7.36 (5H), 7,44 (1H, dd, J=7.0 and 1.3Hz), 7.81 (1H, s), 8.07 (2H,s)

REFERENCE EXAMPLE 85 Preparation of ethyl3-(5-cyanobenzo[b]thien-2-yl)-2-[4-[2-yl)-pyrrolin-2-yl)aminoethoxy]phenyl]propionatehydrochloride

With stirring, 1.3 g of ethyl2-[4-(2-tertbutoxycarbonylaminoethoxy)phenyl]-3-(5-cyanobenzo[b]thien-2-yl)propionatewas dissolved in 50 ml of ethanol, and then stirred. To this solutionwas added 25 ml of ethanol containing 13% (w/v) of hydrochloric acid.The thus prepared mixture was stirred at 50° C. for 30 minutes. Afterdistilling off the solvent, the resulting residue was dissolved in 50 mlof ethanol, and then stirred. The thus prepared solution was mixed with782 mg of 2-ethoxy-1-pyrroline, and the mixture was refluxed underheating for 1.5 hours. After cooling, crystals thus precipitated werecollected by filtration to obtain 1.1 g of the title compound.

mp: 212°-215° C.

¹ H-NMR (CDCl₃) δ: 1.14 (1.5H, t, J=7.0Hz), 1.16 (1.5H, t, J=7.0Hz),2.16 (2H, t, J=7.5Hz), 2.87 (2H, t, J=8.0Hz), 3.20-4.40 (9H), 6.87 (2H,d, J=8.3Hz), 7.13 (1H, s), 7.27 (2H, d, J=8.3Hz), 7.48 (1H, dd, J=8.3and 1.3Hz), 7.92 (1H, d, J=8.3Hz), 8.04 (1H, s), 10.04 (1H, br), 10.40(1H, br)

REFERENCE EXAMPLE 86 Preparation of methyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-indolyl)propionate

In a solvent mixture of 50 ml of tetrahydrofuran and 50 ml of methanolwere dissolved 5.0 g of (5-cyano-2-indolyl)methyltriphenylphosphoniumbromide and 3.6 g of methyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-2-oxoacetate.With stirring at room temperature, 1.07 g of1,8-diazabicyclo[5.4.0]-7-undecene was added to the thus preparedsolution, and the mixture was stirred at the same temperature for 2hours. After distilling off the solvent, the resulting residue waspurified by subjecting it to silica gel column chromatography using adichloromethane/acetone mixture as an eluant, thereby obtaining methyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-indolyl)acrylateas a mixture of E and Z forms. The thus obtained compound was dissolvedin a solvent mixture of 50 ml of tetrahydrofuran and 50 ml of methanol,and the resulting solution was mixed with 5.0 g of palladium oxide.1H₂O.barium sulfate, and then subjected to catalytic hydrogenation undernormal pressure. After removing the catalyst by filtration anddistilling off the solvent, the resulting residue was purified bysubjecting it to silica gel column chromatography using adichloromethane/acetone mixture as an eluant. In this way, 3.5 g of thetitle compound was obtained in a viscous and oily form.

¹ H-NMR (CDCl₃) δ:1.46 (9H, s), 2.00-2.20 (2H, m), 2.95-4.22 (7H, m),4.75-4.90 (1H, br), 6.23 (1H, d), 6.80 (2H, d), 7.18 (2H, d), 7.20-7.40(2H, m), 7.80 (1H, s), 8.80 (1H, br)

The following compounds of Reference Examples 87 to 92 were prepared inaccordance with the procedure described in Reference Example 86.

REFERENCE EXAMPLE 87 methyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyano-2-indolyl)propionate

viscous oil

¹ H-NMR (CDCl₃) δ:1.50 (9H, s), 2.00-2.25 (2H, br), 3.13 (1H, dd),3.37-3.75 (3H, m), 3.97 (1H, dd), 4.70-4.90 (1H, br), 6.37 (1H, s), 6.80(2H, d), 7.10-7.70 (5H, m), 9.25 (1H)

REFERENCE EXAMPLE 88 methyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyano-1-methyl-2-indolyl)propionate

viscous oil

¹ H-NMR (CDCl₃) δ:1.45 (9H, s), 2.00-2.25 (2H, br), 3.13 (1H, dd), 3.60(3H, s), 3.62 (3H, s), 3.90-4.10 (1H, dd), 4.75-4.90 (1H, br), 6.30 (1H,s), 6.80 (2H, d), 7.10-7.70 (5H, m), 9.25 (1H)

REFERENCE EXAMPLE 89 methyl2-[4-[((3R)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyano-1-ethyl-2-indoly)propionate

viscous oil

¹ H-NMR (CDCl₃) δ:1.34 (3H, t, J=7.2Hz), 1.47 (9H, s), 2.00-2.30 (2H,m), 2.90-3.30 (3H, m), 3.40-3.80 (4H, m), 3.66 (3H, s), 4.15 (2H, q,J=7.2Hz), 4.70-5.00 (1H, br), 6.30 (1H, s), 6.84 (2H, d, J=8.8Hz), 7.27(2H, d, J=8.8Hz), 7.26 (1H, d, J=7.0Hz), 7.54 (1H, d, J=7.0Hz)

REFERENCE EXAMPLE 90 methyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyano-1-ethyl-2-indolyl)propionate

viscous oil

¹ H-NMR (CDCl₃) δ:1.32 (3H, t, J=7.2Hz), 1.48 (9H, s), 2.00-2.30 (2H,m), 2.90-3.30 (3H, m), 3.40-3.80 (4H, m), 3.64 (3H, s), 4.15 (2H, q,J=7.2Hz), 4.70-5.00 (1H, br), 6.30 (1H, s), 6.84 (2H, d, J=8.8Hz), 7.26(2H, d, J=8.8Hz), 7.26 (1H, d, J=7.0Hz), 7.54 (1H, d, J=7.0Hz)

REFERENCE EXAMPLE 91 methyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-[1-(2-chloroethyl)-6-cyano-2-indolyl]-propiate

viscous oil

¹ H-NMR (CDCl₃) δ:1.47 (9H, s), 2.00-2.30 (2H, m), 3.00-4.20 (9H, m),3.66 (3H, s), 4.20-4.60 (2H, m), 4.80-5.00 (1H, m), 6.37 (1H, s), 6.84(2H, d), 7.20-7.80 (5H, m)

REFERENCE EXAMPLE 92 methyl2-[4-[(1-tert-butoxycarbonyl-4-piperidinyl)oxy]phenyl]-3-(6-cyano-1-ethyl-2-indolyl)propionate

¹ H-NMR (CDCl₃) δ:1.34 (3H, t), 1.60-2.00 (4H, m), 3.10 (1H, dd),3.30-3.40 (2H, m), 3.57 (1H, dd), 3.62-3.75 (2H, m), 3.90-4.30 (3H, m),4.35 (1H, m), 6.30 (1H, s), 6.90 (2H, d), 7.30 (3H, m), 7.54 (1H, d),7.58 (1H, s)

REFERENCE EXAMPLE 93 Preparation of methyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-1-methyl-2-indolyl)propionate

3.0 g of methyl2-[4-[((3S)-1-tert-butoxy-carbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-indolyl)propionatewas dissolved in 30 ml of N,N-dimethylformamide, and then stirred underice cooling. 270 mg of 60% sodium hydride was added to the abovesolution, and the stirring was continued for 10 minutes. The resultingreaction solution was mixed with 0.4 ml of methyl iodide, and themixture was stirred at room temperature for 1 hour. The thus treatedreaction solution was diluted with a toluene/ethyl acetate mixture, andthen washed with an aqueous solution of ammonium chloride. After dryingthe resulting organic layer to distill off the solvent, the residue thusobtained was purified by subjecting it to silica gel columnchromatography using a dichloromethane/acetone mixture as an elutionsolvent. In this way, 2.0 g of the title compound was obtained in aviscous and oily form.

¹ H-NMR (CDCl₃) δ:1.45 (9H, s), 2.00-2.22 (2H, m), 3.05 (1H, dd),3.35-3.80 (5H, m), 3.63 (3H, s), 4.00 (1H, dd), 4.75-5.00 (1H, br), 6.25(1H, d), 6.85 (2H, d), 7.20-7.50 (2H, m), 7.90 (1H, s)

REFERENCE EXAMPLE 94 Preparation of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidiny)oxy]phenyl]-3-(6-cyano-1,2,3,4-tetrahydro-2-naphthyl)propionate

9.0 g of(6-cyano-1,2,3,4-tetrahydro-2-naphthyl)methyltriphenylphosphoniump-toluenesulfonate was suspended in 150 ml of tetrahydrofuran, followedby gradual addition of 600 mg of 60% sodium hydride. The thus preparedmixture was refluxed under heating for 20 minutes. After cooling down toroom temperature, to the resulting reaction solution was added 10 ml ofa tetrahydrofuran solution containing 4.16 g of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-2-oxoacetate.The resulting reaction mixture was stirred for 10 minutes, and thenrefluxed under heating for 2 hours. After cooling down to roomtemperature, the reaction product thus obtained was dissolved in ethylacetate, and washed with water and saturated sodium chloride aqueoussolution in that order. After drying the resulting organic layer todistill off the solvent, the thus obtained residue was purified bysubjecting it to silica gel column chromatography using a n-hexane/ethylacetate mixture as an elution solvent, thereby obtaining 3.90 g of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]3-(6-cyano-1,2,3,4-tetrahydro-2-naphthyl)acrylatein a yellow oily form as a mixture of E and Z forms. 2.58 g of the thusobtained E/Z mixture was dissolved in 40 ml of ethanol, and theresulting solution was mixed with 650 mg of palladium oxide.1H₂ O.bariumsulfate, and then subjected to catalytic hydrogenation under normalpressure for 5 hours. After removing the catalyst by filtration anddistilling off the solvent, the resulting residue was purified bysubjecting it to silica gel column chromatography using a n-hexane/ethylacetate mixture as an elution solvent. In this way, 1.69 g of the titlecompound was obtained in a yellow oily form.

¹ H-NMR (CDCl₃) δ:1.20 (3H, t, J=7.0Hz), 1.45 (9H, s), 1.50-3.90 (16H,m), 4.10 (2H, q),. 4.82 (1H, m), 6.81 (2H, q, J=9.0Hz), 7.00-7.40 (5H,m)

REFERENCE EXAMPLE 95

Preparation of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-benzimidazolyl)propionate

a) 3.42 g of 3,4-diaminobenzonitrile and 4.06 g of ethylchloroacetoimidate hydrochloride were dissolved in 100 ml of ethanol,and the solution was refluxed under heating for 3 hours. After coolingand distilling off the solvent, the resulting residue was dissolved inethyl acetate, washed with water, and then dried. After distilling offthe solvent, the crystals thus precipitated were collected by filtrationto obtain 2.7 g of 2-chloromethyl-5-benzimidazolecarbonitrile.

mp: 144°-146° C.

¹ H-NMR (CDCl₃) δ:4.83 (2H, s), 7.48 (1H, d, J=7.1Hz), 7.57 (1H, d,J=7.1Hz), 7.95 (1H, s)

b) 1.0 g of 2-chloromethyl-5-benzimidazolecarbonitrile obtained in theabove step a) and 2.19 g of triphenylphosphine were dissolved in 30 mlof 1,2-dichloroethane, and the solution was heated at a temperature of140° C. for 1 hour. After cooling and distilling off the solvent, thethus obtained residue and 2.03 g of ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-2-oxoacetatewere dissolved in a solvent mixture of 20 ml of tetrahydrofuran and 20ml of ethanol. With stirring at room temperature, 1.1 g of1,8-diazabicyclo[5.4.0]-7-undecene was added to the thus preparedsolution, and the mixture was stirred at the same temperature for 72hours. After distilling off the solvent, the resulting residue waspurified by subjecting it to silica gel column chromatography using achloroform/ethanol mixture as an elution solvent, thereby obtaining 1.5g of oily ethyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-benzimidazolyl)acrylateas a mixture of E and Z forms. The thus obtained E/Z mixture wasdissolved in a solvent mixture consisting of 50 ml of tetrahydrofuranand 50 ml of ethanol, and the resulting solution was mixed with 1.5 g ofpalladium oxide.1H₂ O.barium sulfate, and then subjected to catalytichydrogenation under normal pressure. After removing the catalyst byfiltration and distilling off the solvent, the resulting residue waspurified by subjecting it to silica gel column chromatography using achloroform/ethanol mixture as an elution solvent. In this way, 320 mg ofthe title compound was obtained in a viscous and oily form.

¹ H-NMR (CDCl₃) δ:1.14 (3H, t, J=7.0Hz), 1.48 (9H, s), 1.90-2.30 (2H,br), 3.05-3.90 (6H, m), 4.12 (2H, q, J=7.0Hz), 4.00-4.30 (1H), 4.70-4.95(1H, br), 6.79 (2H, d, J =8.8Hz), 7.19 (2H, d, J=8.8Hz), 7.35-8.10 (3H,m)

FD MS (m/z): 504 (M⁺), 505 (M⁺ +1)

REFERENCE EXAMPLE 96 Preparation(+)-((2S)-1-p-toluenesulfonyl-2-pyrrolidinyl)methyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyano-2-indoyl)propionateand (-)-[((2S)-1-p-toluenesulfonyl-2-2-pyrrolidinyl)methyl2-[4[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl]oxy]phenyl]-3-(6-cyano-2-indolyl)propionate

a) An aqueous solution containing 3 g of sodium hydroxide dissolved in10 ml of water was added to 100 ml of a methanol solution containing 22g of methyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyano-2-indolyl)propionate,and the resulting mixture was stirred at room temperature for 24 hours.After distilling off the solvent, the remaining portion was adjusted topH 4-5 with citric acid and then extracted with ethyl acetate. By dryingthe extract to distill off the solvent, 20 g of2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyano-2-indolyl)propionicacid was obtained.

IR (KBr): 3352, 2218, 1710, 1677 cm⁻¹

b) In 300 ml of dioxane were dissolved 20 g of2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyano-2-indolyl)propionicacid obtained in the above step a) and 11.9 g of((2S)-1-p-toluenesulfonyl-2pyrrolidinyl)methanol. The thus preparedsolution was mixed with a catalytically effective amount of4-dimethylaminopyridine and 9 g of 1,3-dicyclohexylcarbodiimide whilestirring under ice cooling, and the mixture was stirred at roomtemperature for 24 hours. After removing precipitated materials byfiltration and distilling off the solvent, the resulting residue waspurified by subjecting it to silica gel column chromatography using achloroform/acetone mixture as an elution solvent, thereby obtaining 10.5g of (+)-((2S)-1-p-toluenesulfonyl-2-pyrrolidinyl)methyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyano-2-indolyl)propionate.

¹ H-NMR (CDCl₃) δ:1.00-1.80 (4H, m), 1.46 (9H, s), 2.00-2.30 (2H, m),2.43 (3H, s), 3.00-4.40 (12H, m), 4.75-5.00 (1H, m), 6.30 (1H, s), 6.82(2H, d), 7.10-7.90 (9H, m), 9.00 (1H, s)

By eluting the column again with the same solvent system, 9.5 g of(-)-((2S)-1-p-toluenesulfonyl-2-pyrrolidinyl)methyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyano-2-indolyl)propionatewas obtained.

¹ H-NMR (CDCl₃) δ: 1.10-2.00 (4H, m), 1.44 (9H, s), 2.00-2.25 (2H, m),2.41 (3H, s), 2.95-4.10 (10H, m), 4.20 (2H, d), 4.70-4.90 (1H, m), 6.25(1H, s), 6.80 (2H, d), 7.10-7.80 (9H, m), 9.20 (1H, s)

REFERENCE EXAMPLE 97 Preparation of methyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]3-(6-cyano-1-ethoxycarbonylmethyl-2-indolyl)propionate

3.0 g of methyl2-[4-[((3S)-1-tert-butoxy-carbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyano-2-indolyl)propionatewas dissolved in 30 ml of N,N-dimethylformamide. The thus preparedsolution was mixed with 280 mg of 60% sodium hydride while stirringunder ice cooling, and the stirring was continued for 20 minutes at thesame temperature. The resulting reaction solution was mixed with 0.7 mlof bromoacetate, and the mixture was stirred for 1 hour. The thustreated reaction solution was mixed with dilute hydrochloric acid,extracted with a toluene/ethyl acetate mixture, washed with water, andthen dried. After distilling off the solvent, the resulting residue waspurified by subjecting it to silica gel column chromatography using adichloromethane/acetone mixture as an elution solvent. In this way, 3.2g of the title compound was obtained in a viscous and oily form.

¹ H-NMR (CDCl₃) δ:1.26 (3H, t), 1.46 (9H, s), 3.02 (1H, dd), 3.30-3.70(5H, m), 3.66 (3H, s), 4.00 (1H, dd), 4.20 (2H, q), 4.80 (2H, s),4.78-4.90(1H, m), 6.40 (1H, s), 6.90 (2H, d), 7.20-7.70 (5H, m)

REFERENCE EXAMPLE 98 Preparation of2-[4-[(3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyano-2-indolyl)propanol

2.7 g of methyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyano-2-indolyl)propionatewas dissolved in 30 ml of tetrahydrofuran, followed by the addition of660 mg of sodium borohydride. 12 ml of methanol was added dropwise tothe thus prepared solution while stirring under ice cooling, and theresulting mixture was stirred at room temperature for 3 hours. Theresulting reaction solution was mixed with 10% citric acid aqueoussolution, extracted with dichloromethane, and then dried. Afterdistilling off the solvent, the resulting residue was purified bysubjecting it to silica gel Column chromatography using adichloromethane/methanol mixture as an elution solvent. In this way, 2.2g of the title compound was obtained in an oily form.

¹ H-NMR (CDCl₃) δ:1.48 (9H, s), 1.95-2.25 (2H, m), 2.48 (1H, t),3.00-3.22 (2H, m), 3.40-3.69 (6H, m), 3.70-3.90 (1H, m), 4.70-4.90 (1H,m), 6.21 (1H, s), 6.80 (2H, d), 7.00-7.65 (5H, m), 9.20 (1H, s)

REFERENCE EXAMPLE 99 Preparation of ethyl2-[2-[4[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-hydroxypropyl]-6-cyano-1-indoleacetate

2.0 g of2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyano-2-indolyl)propanolwas dissolved in 30 ml of N,N-dimethylformamide. 280 mg of 60% sodiumhydride was added to the above solution while stirring under icecooling, and the stirring was continued for 20 minutes at the sametemperature. The resulting reaction solution was mixed with 0.5 ml ofethyl bromoacetate, and the mixture was stirred for 1 hour. The thustreated reaction solution was mixed with an aqueous solution of ammoniumchloride, extracted with a toluene/ethyl acetate mixture, washed withwater, and then dried. After distilling off the solvent, the resultingresidue was purified by subjecting it to silica gel columnchromatography using a dichloromethane/acetone mixture as an elutionsolvent. In this way, 1.5 g of the title compound was obtained in aviscous and oily form.

¹ H-NMR (CDCl₃) δ:1.23 (3H, t), 1.45 (9H, s), 1.90-2.20 (2H, s), 4.20(2H, q), 4.50-4.90 (3H), 6.20 (1H, s), 6.78 (2H, d)

REFERENCE EXAMPLE 100 Preparation of2-[2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]ethyl]-6-indolecarbonitrile

a) In 40 ml of tetrahydrofuran were dissolved 1.31 g ofp-hydroxybenzaldehyde, 1.87 g of(3R)-1-tert-butoxycarbonyl-3-hydroxypyrrolidine and 2.88 g oftriphenylphosphine. With stirring at room temperature, 1.91 g of diethylazodicarboxylate was added to the thus prepared solution, and themixture was stirred for 45 minutes. After distilling off the solvent,the resulting residue was purified by subjecting it to silica gel columnchromatography using a benzene/ethyl acetate mixture as an elutionsolvent. In this way, 2.9 g of4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]benzaldehyde wasobtained in an oily form.

¹ H-NMR (CDCl₃) δ:1.48 (9H, s), 2.00-2.40 (2H, m), 3.30-3.80 (4H, m),4.90-5.10 (1H, m), 6.98 (2H, d, J=9.0Hz), 7.84 (2H, d, J=9.0Hz), 9.89(1H, s)

b) 0.93 g of4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]benzaldehyde obtainedin the above step a) and 1.6 g of(6-cyano-2-indolyl)methyltriphenylphosphonium bromide were dissolved ina solvent mixture of 20 ml of methanol and 20 ml of tetrahydrofuran. 490mg of 1,8-diazabicyclo[5.4.0]-7-undecene was dissolved in the thusprepared solution while stirring under ice cooling, and the resultingmixture was stirred at room temperature for 3 hours. After distillingoff the solvent, the resulting residue was purified by subjecting it tosilica gel column chromatography using a chloroform/methanol mixture asan elution solvent, thereby obtaining 700 mg of2-[2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]vinyl]-6-indolecarbonitrileas a mixture of E and Z forms.

¹ H-NMR (CDCl₃) δ:1.43 (9H, s), 1.90-2.23 (2H, m), 3.30-3.70 (4H, m),4.75-4.95 (1H, m), 8.65 (1H, br)

c) 700 mg of2-[2-[4-[((3S)-1-tert-butoxy-carbonyl-3-pyrrolidinyl)oxy]phenyl]vinyl]-6-indolecarbonitrileobtained in the above step b) was dissolved in a solvent mixture of 20ml of methanol and 40 ml of tetrahydrofuran. 70 mg of palladiumoxide.1H₂ O.barium sulfate was added to the solution prepared above, andthe mixture was subjected to catalytic hydrogenation under normalpressure for 3 hours. After removing the catalyst by filtration anddistilling off the solvent, the resulting residue was purified bysubjecting it to silica gel column chromatography using achloroform/methanol mixture as an elution solvent. In this way, 650 mgof the title compound was obtained in a viscous and oily form.

¹ H-NMR (CDCl₃) δ:1.50 (9H, s), 1.95-2.20 (2H, m), 4.70-4.90 (1H, m),6.30 (1H, s), 6.75 (2H, d), 7.10 (2H, d), 7.10-7.65 (3H, m), 9.46 (1H,br)

REFERENCE EXAMPLE 101 Preparation of ethyl2-[2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]ethyl]-6-cyano-1-indoleacetate

2.4 g of2-[2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]ethyl]-6-indolecarbonitrilewas dissolved in 50 ml of N,N-dimethylformamide. 300 mg of 60% sodiumhydride was added to the thus prepared solution during stirring underice cooling, and the resulting mixture was warmed up to room temperatureand stirred at the same temperature for 20 minutes. 0.76 ml of ethylbromoacetate was added to the above mixture during stirring under icecooling, followed by stirring for 1 hour. The resulting reactionsolution was mixed with an ammonium chloride aqueous solution andextracted with a toluene/ethyl acetate mixture, and the resultingorganic layer was washed with water and dried. After distilling off thesolvent, the resulting residue was purified by subjecting it to silicagel column chromatography using a dichloromethane/acetone mixture as anelution solvent. In this way, 2.3 g of the title compound was obtainedin a viscous and oily form.

¹ H-NMR (CDCl₃) δ:1.2 (3H, t), 2.00-2.20 (2H, m), 2.95 (4H, s),3.30-3.60 (4H, m), 4.18 (2H, q), 4.70 (2H, s), 6.36 (1H, s), 6.75 (2H,d), 7.00-7.60 (5H, m)

REFERENCE EXAMPLE 102 Preparation of2-[[4-[(1-tert-butoxycarbonyl-4-piperidinyl)oxy]phenyl]methyl]-5-benzofurancarbonitrile

a) 5.07 g of potassium hydroxide was added to 30 ml of diethyleneglycol, and the mixture was stirred at room temperature during which 5.5g of 80% hydrazine dihydrate.2H₂ O and 5.0 g of5-bromo-2-(4-methoxybenzoyl)benzofuran were further added. The thusprepared mixture was refluxed under heating. After cooling, theresulting reaction solution was adjusted to pH 4-5, extracted withbenzene, and then dried. After distilling off the solvent, the resultingresidue was purified by subjecting it to silica gel columnchromatography using a n-hexane/isopropanol mixture as an elutionsolvent, thereby obtaining 3.95 g of5-bromo-2-(4-methoxybenzyl)benzofuran as a brown oily material.

¹ H-NMR (CDCl₃) δ:3.80 (3H, s), 4.02 (2H, s), 6.23 (1H, s), 6.90 (2H, d,J=9.0Hz), 7.20-7.40 (4H, m), 7.57 (1H, m)

b) 3.95 g of 5-bromo-2-(4-methoxybenzyl)benzofuran obtained in the abovestep a) and 1.67 g of cuprous cyanide were suspended in 20 ml ofN-methyl-2-pyrrolidone, and the suspension was heated at a temperatureof 200° to 220° C. in a stream of nitrogen. After cooling, the resultingreaction product was dissolved in chloroform, and insoluble materialswere removed by filtration. The resulting organic layer was washed withwater and concentrated to dryness. Thereafter, the thus obtained residuewas purified by subjecting it to silica gel column chromatography usinga n-hexane/iso-propyl ether mixture as an elution solvent, therebyobtaining 3.10 g of 2-(4-methoxybenzyl)-5-benzofurancarbonitrile.

mp: 78°-80° C.

¹ H-NMR (CDCl₃) δ: 3.80 (3H, s), 4.10 (2H, s), 6.39 (1H, s), 6.90 (2H,d, J=9.0Hz), 7.22 (2H, d), 7.46 (2H), 7.78 (1H, s)

c) 3.0 g of 2-(4-methoxybenzyl)-5-benzofurancarbonitrile obtained in theabove step b) was dissolved in 30 ml of dichloromethane, and thesolution was cooled down to -50° C. With stirring, to the above solutionwas added dropwise 20 ml of a dichloromethane solution containing 2.23ml of boron tribromide. After gradually warming up the mixture to roomtemperature, stirring was continued for 1 hour. The resulting reactionsolution was diluted with chloroform, washed with dilute hydrochloricacid, and then dried. After distilling off the solvent, the crystalsthus precipitated were collected by filtration to obtain 2.48 g of2-(4-hydroxybenzyl)-5-benzofurancarbonitrile in the form of yellow prismcrystals.

¹ H-NMR (CDCl₃) δ:4.02 (2H, s), 6.45 (1H, s), 6.77 (2H, d, J=9.0Hz),7.10 (2H, d), 7.48 (2H), 7.81 (1H, s)

d) In 50 ml of tetrahydrofuran were dissolved 1.50 g of2-(4-hydroxybenzyl)-5-benzofurancarbonitrile obtained in the above stepc), 2.37 g of triphenylphosphine and 1.21 g of1-tert-butoxycarbonyl-4-hydroxypiperidine. With stirring at roomtemperature, the thus prepared solution was mixed with 1.57 g of diethylazodicarboxylate, and the stirring was continued for 40 hours. Afterdistilling off the solvent, the resulting residue was purified bysubjecting it to silica gel column chromatography using a n-hexane/ethylacetate mixture as an elution solvent. In this way, 1.30 g of the titlecompound was obtained in the form of colorless needle crystals.

mp: 144°-146° C.

¹ H-NMR (CDCl₃) δ:1.47 (9H, s), 1.60-2.00 (4H, m), 3.20-3.90 (4H, m),4.05 (2H, s), 4.44 (1H, m), 6.41 (1H, s), 6.87 (2H, d, J=9.0Hz), 7.20(2H, d, J=9.0Hz), 7.47 (2H), 7.79 (1H, s)

REFERENCE EXAMPLE 103 Preparation of3-[3-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]propyl]-5-benzofurancarbonitrile

a) 2.14 g of (5-cyano-3-benzofuranyl)methyltriphenylphosphonium chlorideand 0.7 g of 4-methoxyphenylacetaldehyde were dissolved in a solventmixture of 100 ml of tetrahydrofuran and 100 ml of ethanol. The thusprepared solution was mixed with 0.71 g of1,8-diaza-bicyclo[5.4.0]-7-undecene and stirred for 24 hours. Afterdistilling off the solvent, the resulting residue was purified bysubjecting it to silica gel column chromatography using toluene as anelution solvent, thereby obtaining 0.86 g of yellow and oily3-[3-(4-methoxyphenyl)allyl]-5-benzofurancarbonitrile as a mixture of Eand Z forms. The thus obtained E/Z mixture was dissolved in 100 ml ofethanol, and the solution was subjected to catalytic hydrogenation undernormal pressure in the presence of 370 mg of 5% palladium carboncatalyst. Thereafter, the catalyst was removed by filtration, and thesolvent was distilled off to obtain 0.6 g of3-[3-(4-methoxyphenyl)propyl]-5-benzofurancarbonitrile. The thusobtained methoxy compound was dissolved in 20 ml of dichloromethane.With stirring at -40° C., to the resulting solution was added dropwise10 ml of a dichloromethane solution containing 0.4 ml of borontribromide. The resulting mixture was warmed up to room temperature andstirred for 2 hours. The resulting reaction solution was poured intocold dilute hydrochloric acid and extracted with chloroform. Afterdrying the resulting organic layer and distilling off the solvent, thethus obtained residue was purified by subjecting it to silica gel columnchromatography using toluene as an elution solvent. In this way, 280 mgof 3-[3-(4-hydroxyphenyl)propyl]-5-benzofurancarbonitrile.

¹ H-NMR (CDCl₃) δ:2.0 (2H, m), 2.64 (4H, m), 6.80 (2H, d), 7.16 (2H, d),7.52 (3H, m), 7.79 (1H)

b) In 20 ml of tetrahydrofuran were dissolved 280 mg of3-[3-(4-hydroxyphenyl)propyl]-5-benzofurancarbonitrile obtained in theabove step a), 280 mg of (3R)-1-tert-butoxycarbonyl-3-hydroxypyrrolidineand 400 mg of triphenylphosphine. With stirring at room temperature, thethus prepared solution was mixed with 265 mg of diethylazodicarboxylate, and the stirring was continued for 24 hours. Afterdistilling off the solvent, the resulting residue was purified bysubjecting it to silica gel column chromatography using a n-hexane/ethylacetate mixture as an elution solvent. In this way, 400 mg of the titlecompound was obtained in a yellow and oily form.

¹ H-NMR (CDCl₃) δ:1.46 (9H, s), 2.05 (4H, m), 2.66 (4H, m), 3.60 (4H,br), 4.85 (1H, br), 6.85 (2H, d), 7.05 (2H, d), 7.53 (3H, m), 7.83 (1H)

REFERENCE EXAMPLE 104 Preparation of4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]-3-methoxybenzaldehyde

In 50 ml of tetrahydrofuran were dissolved 3.04 g of vanillin, 3.74 g of(3R)-1-tert-butoxycarbonyl-3-hydroxypyrrolidine and 5.24 g oftriphenylphosphine. The thus prepared solution was mixed with 4.00 g ofdiethyl azodicarboxylate, and the mixture was stirred at roomtemperature for 18 hours. After concentrating the resulting reactionsolution, the thus obtained residue was purified by subjecting it tosilica gel column chromatography using an ethanol/chloroform mixture asan elution solvent. In this way, 5.0 g of the title compound wasobtained in an oily form.

¹ H-NMR (CDCl₃) δ: 1.47 (9H, s), 2.00-2.40 (2H, m), 3.50-3.80 (4H, m),3.90 (3H, s), 5.02 (1H, br), 6.80-7.60 (3H, m), 9.86 (1H, s)

REFERENCE EXAMPLE 105 Preparation of4-[[4-(N-acetyl)aminomethylcyclohexyl]methoxy]benzaldehyde

The title compound was prepared in accordance with the procedure asdescribed in Reference Example 104.

¹ H-NMR (CDCl₃) δ: 0.80-2.10 (10H, m), 3.13 (2H, dd, J=6.1 and 6.1Hz),3.84 (2H, d, J=6.1Hz), 5.56 (1H, br), 6.97 (2H, d, J=8.7Hz), 7.82 (2H,d, J=8.7Hz), 9.88 (1H, s)

REFERENCE EXAMPLE 106 Preparation of3-acetoxy-4-[((3S)-1-acetyl-3-pyrrolidinyl)oxy]benzaldehyde

a) 5.5 g of4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]-3-methoxybenzaldehydewas dissolved in 20 ml of dichloromethane, followed by the addition of30 ml of formic acid. The thus prepared mixture was stirred at roomtemperature for 1 hour and then at 50° C. for 1 hour. The solvent andformic acid were distilled off under a reduced pressure, and theresulting residue was dissolved in 100 ml of tetrahydrofuran. The thusprepared solution was mixed with 2.68 g of acetyl chloride, and 8.63 gof triethylamine was added dropwise thereto while stirring under icecooling. Thereafter, the solvent was distilled off, and the residue thusobtained was dissolved in chloroform, and washed with water, followed bydrying of the resulting organic layer. After distilling off the solvent,the resulting residue was purified by subjecting it to silica gel columnchromatography using toluene as an elution solvent, thereby obtaining4.3 g of 4-[((3S)-1-acetyl-3-pyrrolidinyl)oxy]-3-methoxybenzaldehyde asa viscous oil.

¹ H-NMR (CDCl₃) δ:2.04 (5H, m), 3.50-4.00 (4H, m), 3.90 (3H, s), 5.10(1H, br), 6.80-7.60 (3H, m), 9.86 (1H, s)

b) 4.3 g of 4-[((3S)-1-acetyl-3-pyrrolidinyl)oxy]-3-methoxybenzaldehydeobtained in the above step a) was dissolved in 40 ml of dichloromethane,and the solution was cooled down to -70° C. With stirring, to the abovesolution was added dropwise 12.6 g of boron tribromide. The thusprepared reaction solution was warmed up to 0° C. and then poured intoice water, followed by extraction with chloroform. The resulting organiclayer was concentrated to dryness, and the thus obtained residue wascrystallized from a chloroform/n-hexane solvent system, therebyobtaining 2.0 g of4-[((3S)-1-acetyl-3-pyrrolidinyl)oxy]-3-hydroxybenzaldehyde. The thusobtained compound was used in the following reaction without furtherpurification.

¹ H-NMR (CDCl₃ :DMSO-d₆ =9:1) δ:2.00-2.50 (5H, m), 3.50-4.00 (4H, m),5.32 (1H, br), 6.96 (1H, d, J=8.0Hz), 7.22-7.58 (2H, m), 9.81 (1H, s)

c) 1.1 g of 4-[((3S)-1-acetyl-3-pyrrolidinyl)oxy]-3-hydroxybenzaldehydeobtained in the above step b) was suspended in 5 ml of pyridine,followed by the addition of 0.86 g of acetic anhydride. The thusprepared mixture was stirred at room temperature for 1 hour. Afterdrying the resulting reaction solution under a reduced pressure, thethus obtained residue was purified by subjecting it to silica gel columnchromatography using an ethanol/chloroform mixture as an elutionsolvent. In this way, 1.30 g of the title compound was obtained in anoily form.

¹ H-NMR (CDCl₃) δ:2.06 (3H, s), 2.28 (3H, s), 2.00-2.40 (2H, m),3.40-3.90 (4H, m), 5.60 (1H, br), 6.90-7.90 (3H, m), 9.90 (1H, s)

REFERENCE EXAMPLE 107 Preparation of4-[(1-trityl-4-imidazolyl)methoxy]benzaldehyde

1.22 g of p-hydroxybenzaldehyde and 4.08 g of4-chloromethyl-1-tritylimidazole were dissolved in 40 ml ofN,N-dimethylformamide, followed by adding 1.66 g of anhydrous potassiumcarbonate and subsequently stirring at room temperature for 40 hours.The thus prepared reaction solution were partitioned between water andbenzene, and the resulting organic layer was concentrated to dryness.The resulting residue was purified by subjecting it to silica gel columnchromatography using chloroform as an elution solvent, and the thuspurified product was crystallized from a n-hexane/benzene solventsystem. In this way, 2.3 g of the title compound was obtained.

mp: 181°-182° C.

¹ H-NMR (CDCl₃) δ:5.09 (2H, s), 6.90 (1H, d, J=1.1Hz), 7.00-7.40 (17H,m), 7.47 (1H, d, J=1.1Hz), 7.81 (2H, d, J=8.8Hz), 9.88 (1H, s)

REFERENCE EXAMPLE 108 Preparation of2-[2-[4-[((3S)-1-acetyl-3-pyrrolidinyl)oxy]-3-hydroxyphenyl]ethyl]-5-furancarbonitrile

a) 1.3 g of3-acetyloxy-4-[((3S)-1-acetyl-3-pyrrolidinyl)oxy]benzaldehyde and 2.22 gof (5-cyano-2-benzofuranyl)methyltriphenylphosphonium chloride weredissolved in a solvent mixture of 10 ml of tetrahydrofuran and 10 ml ofethanol, followed by the addition of 0.943 g of1,8-diazabicyclo[5.4.0]-7-undecene and by subsequent stirring at roomtemperature for 2 hours. To the mixture obtained were further added 1.88g of 1,8-diazabicyclo[5.4.0]-7-undecene and 3 ml of water, followed bystirring at room temperature for 2 hours. The thus prepared reactionsolution was adjusted to pH 4-5 with 10% citric acid aqueous solutionand concentrated under a reduced pressure, and the resulting residue wasextracted with chloroform, and then dried. After distilling off thesolvent, the resulting residue was purified by subjecting it to silicagel column chromatography using a chloroform/ethanol mixture as anelution solvent, thereby obtaining 1.8 g of2-[2-[4-[((3S)-1-acetyl-3-pyrrolidinyl)oxy]-3-hydroxyphenyl]vinyl]-5-benzofurancarbonitrileas a powdery mixture of E and Z forms.

¹ H-NMR (CDCl₃) δ:2.00-2.50 (5H, m), 3.40-4.00 (4H, m), 5.00 (1H, br),6.30-7.90 (9H, m)

b) 1.8 g of2-[2-[4-[((3S)-1-acetyl-3-pyrrolidinyl)oxy]-3-hydroxyphenyl]vinyl]-5-benzofurancarbonitrileobtained in the above step a) was dissolved in a solvent mixture of 300ml of tetrahydrofuran and 300 ml of ethanol, and the solution wassubjected to catalytic reduction under normal pressure for 5 hours inthe presence of 340 mg of palladium oxide.1H₂ O.barium sulfate. Afterremoving the catalyst by filtration and concentrating the resultingfiltrate, the crystals thus precipitated were collected by filtration toobtain 1.6 g of the title compound in the form of colorless crystals.

mp: 191°-193° C.

IR (KBr): 2224, 1644, 1512 cm⁻¹

¹ H-NMR (CDCl₃) δ:2.09 (3H, s), 2.00-2.50 (2H, m), 3.04 (4H, s),3.50-3.90 (4H, m), 4.96 (1H, br), 6.40 (1H, s), 6.50-6.90 (3H, m), 7.50(2H, s), 7.80 (1H, s)

The following compounds of Reference Examples 109 and 110 were preparedin accordance with the procedure as described in Reference Example 108.

REFERENCE EXAMPLE 1092-[2-[4-[[4-(N-acetyl)aminomethylcyclohexyl]methoxy]phenyl]ethyl]-5-benzofurancarbonitrile

mp: 159°-161° C.

¹ H-NMR (CDCl₃) δ:0.80-2.10 (10H, m), 1.99 (3H, s), 3.04 (4H, s), 3.19(2H, dd, J=6.1 and 6.1Hz), 3.63 (2H, d, J =6.1Hz), 5.50 (1H, br), 6.40(1H, s), 6.79 (2H, d, J=8.3Hz), 7.08 (2H, d, J=8.3Hz), 7.49 (2H), 7.79(1H)

REFERENCE EXAMPLE 1102-[2-[4-[(1-trityl-4-imidazolyl)methoxy]phenyl]ethyl]-5-benzofurancarbonitrile

mp: 165°-167° C.

¹ H-NMR (CDCl₃) δ:3.03 (4H, s), 4.97 (2H, s), 6.38 (1H, s), 6.90-7.70(22H, m), 7.78 (1H, s)

REFERENCE EXAMPLE 111 Preparation of2-[2-[4-[(1-imidazolyl)methyl]phenyl]ethyl]-5-benzofurancarbonitrile

a) 911 mg of 4-hydroxymethylbenzaldehyde and 2.0 g of(5-cyano-2-benzofuranyl)methyltriphenylphosphonium chloride weredissolved in a solvent mixture of 10 ml of tetrahydrofuran and 10 ml ofethanol, followed by the addition of 845 mg of1,8-diazabicyclo[5.4.0]-7-undecene and by subsequent stirring at roomtemperature for 1 hour. After distilling off the solvent, the resultingresidue was purified by subjecting it to silica gel columnchromatography, thereby obtaining 2.3 g of2-[2-(4-hydroxymethylphenyl)vinyl]-5-benzofurancarbonitrile as a mixtureof E and Z forms. 2.3 g of the thus obtained E/Z mixture was dissolvedin a solvent mixture of 10 ml of tetrahydrofuran and 10 ml of ethanol,and the solution was subjected to catalytic hydrogenation under normalpressure for 7 hours in the presence of 800 mg of palladium oxide.1H₂O.barium sulfate. After removing the catalyst by filtration anddistilling off the solvent, the resulting residue was purified bysubjecting it to silica gel column chromatography using chloroform as anelution solvent to obtain 835 mg of2-[2-(4-hydroxymethylphenyl)ethyl]-5-benzofurancarbonitrile as crystals.

mp: 123°-124° C.

¹ H-NMR (CDCl₃) δ:1.60 (1H, s), 3.10 (4H, s), 4.67 (2H, s), 6.41 (1H,s), 7.20 (2H, d, J=8.2Hz), 7.34 (2H, d, J=8.2Hz), 7.52 (2H, s), 7.82(1H, s)

b) 835 mg of 2-[2-(4-hydroxymethylphenyl)ethyl]-5-benzofurancarbonitrileobtained in the above step a) was dissolved in 15 ml-of thionylchloride, and the solution was stirred at room temperature for 1 hour.Thereafter, thionyl chloride was distilled off, and the resultingresidue was dissolved in 30 ml of acetonitrile, together with 550 mg ofN-acetylimidazole and 600 mg of sodium iodide. The thus preparedsolution was refluxed for 3 hours. After distilling off the solvent, theresulting residue was purified by subjecting it to silica gel columnchromatography using chloroform as an elution solvent. In this way, 800mg of the title compound was obtained in the form of brown crystals.

mp: 72°-73° C.

¹ H-NMR (CDCl₃) δ:3.08 (4H, s), 5.09 (2H, s), 6.40 (1H, s), 6.89 (1H,s), 7.00-7.18 (5H, m), 7.49 (2H, s), 7.56 (1H, s), 7.79 (1H, s)

REFERENCE EXAMPLE 112 Preparation of4-[2-(5-cyano-2-benzofuranyl)ethyl]benzoic acid

a) 5.17 g of methyl 4-formylbenzoate and 13.97 g of(5-cyano-2-benzofuranyl)methyltriphenylphosphonium chloride weredissolved in a solvent mixture of 50 ml of tetrahydrofuran and 50 ml ofmethanol. A 5.02 g of 1,8-diazabicyclo[5.4.0]-7-undecene was added tothe thus prepared solution while stirring under ice cooling, followed bystirring at room temperature for 2 hours. By collecting the precipitatedcrystals through a filter, methyl4-[2-(5-cyano-2-benzofuranyl)vinyl]benzoate was obtained as a mixture ofE and Z forms. The thus obtained crystals were dissolved in a solventmixture of 300 ml of tetrahydrofuran and 100 ml of ethanol, and thesolution was subjected to catalytic hydrogenation under normal pressurefor 2 hours in the presence of 2.0 g of palladium oxide.1H₂ O.bariumsulfate. After removing the catalyst by filtration and concentrating theresulting filtrate, the thus obtained residue was purified by subjectingit to silica gel column chromatography using benzene as an elutionsolvent to obtain 8.1 g of methyl4-[2-(5-cyano-2-benzofuranyl)ethyl]benzoate in the form of prismcrystals.

mp: 114°-115° C.

¹ H-NMR (CDCl₃) δ:3.13 (4H, s), 3.90 (3H, s), 6.31 (1H, s), 7.26 (2H, d,J=8.5Hz), 7.50 (2H, s), 7.80 (1H, s), 7.98 (2H, d, J=8.5Hz)

b) 1.5 g of methyl 4-[2-(5-cyano-2-benzofuranyl)ethyl]benzoate obtainedin the above step a) was dissolved in a solvent mixture of 20 ml oftetrahydrofuran and 20 ml of ethanol, followed by the addition of 11 mlof 1N sodium hydroxide aqueous solution and by subsequent stirring atroom temperature for 14 hours. Thereafter, the resulting reactionsolution was adjusted to pH 2 with concentrated hydrochloric acid, andthe thus precipitated crystals were collected by filtration, washed withwater, and then dried. In this way, 1.41 g of the title compound wasobtained.

mp: 234°-235° C.

¹ H-NMR (DMSO-d₆) δ:3.13 (4H, s), 6.70 (1H, s), 7.44 (2H, d, J=8.0Hz),7.69 (2H, s), 7.88 (2H, d, J=8.0Hz), 8.06 (1H, s)

REFERENCE EXAMPLE 113 Preparation of2-[2-[4-[(4-methyl-1-piperazinyl)carbonyl]phenyl]ethyl]-5-benzofurancarbonitrile

1.35 g of 4-[2-(5-cyano-2-benzofuranyl)ethyl]benzoic acid was refluxedfor 2 hours in 15 ml of thionyl chloride. Thereafter, thionyl chloridewas distilled off, and the thus obtained residue was dissolved in 10 mlof tetrahydrofuran. The thus prepared solution was added dropwise to 20ml of a tetrahydrofuran solution containing 1.0 g of 1-methylpiperazinewhich was stirred under ice cooling. After stirring at room temperaturefor 1 hour and subsequent removal of the solvent by distillation, theresulting residue was dissolved in chloroform and washed with saturatedsodium bicarbonate aqueous solution. After drying the resulting organiclayer and distilling off the solvent, the thus obtained residue waspurified by subjecting it to silica gel column chromatography using achloroform/ethanol mixture as an elution solvent. In this way, 1.35 g ofthe title compound was obtained in the form of crystals.

mp: 115°-116° C.

¹ H-NMR (DMSO-d₆) δ:2.34 (3H, s), 2.43 (4H, br), 3.11 (4H, s), 3.64 (4H,br), 6.42 (1H, s), 7.26 (2H, d, J=9.0Hz), 7.40 (2H, d, J=9.0Hz), 7.50(2H, s), 7.80 (1H, s)

REFERENCE EXAMPLE 114

Preparation of2-[2-[4-[[(2-pyrazinyl)amino]carbonyl]phenyl]ethyl]-5-benzofurancarbonitrile

a) At room temperature, 1 g of4-[2-(5-cyano-2-benzofuranyl)ethyl]benzoic acid and 578 mg of1-hydroxybenzotriazole were dissolved in 100 ml of dichloromethane,followed by the addition of 780 mg of 1,3-dicyclohexylcarbodiimide andby subsequent stirring at the same temperature for 3 hours. Afterdistilling off the solvent, the resulting residue was purified bysubjecting it to silica gel column chromatography using achloroform/ethanol mixture as an elution solvent, and the thus purifiedproduct was crystallized from a benzene/n-hexane mixture. In this way,1.1 g of2-[2-[4-[[(1-benzotriazolyl)oxy]carbonyl]phenyl]ethyl]-5-benzofurancarbonitrilewas obtained in a powdery form.

mp: 171°-172° C.

¹ H-NMR (CDCl₃) δ: 3.14 (4H, s), 6.48 (1H, s), 7.30-8.30 (11H, m)

MS (m/z): 409 (M⁺ +1)

b) 100 mg of2-[2-[4-[[(1-benzotriazolyl)oxy]carbonyl]phenyl]ethyl]-5-benzofurancarbonitrileobtained in the above step a) and 23.3 mg of aminopyrazine weredissolved in 2 ml of N,N-dimethylformamide. The thus prepared solutionwas mixed with 13.0 mg of 60% sodium hydride and stirred at roomtemperature for 2 hours. The resulting reaction solution was dilutedwith ethyl acetate, washed with water, and then dried. After distillingoff the solvent, the resulting residue was purified by subjecting it tosilica gel column chromatography to obtain 55 mg of the title compoundin a powdery form.

mp: 183°-185° C.

¹ H-NMR (CDCl₃) δ: 3.17 (4H, s), 6.42 (1H, s), 7.34 (2H, d, J=9.0Hz),7.59 (2H, s), 7.81 (1H, s), 7.90 (2H, d, J=9.0Hz), 8.26 (1H, dd, J=3.0and 1.6Hz), 8.40 (1H, d, J=3.0Hz), 8.65 (1H, br), 9.74 (1H, d, J=1.6Hz)

REFERENCE EXAMPLE 115 Preparation of methyl[5-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]-2-[2-(5-cyano-2-benzofuranyl)ethyl]phenyl]oxyacetate

a) in 40 ml of tetrahydrofuran were dissolved 1.38 g of2,4-dihydroxybenzaldehyde, 1.87 g of(3R)-1-tert-butoxycarbonyl-3-hydroxypyrrolidine and 2.88 g oftriphenylphosphone. The thus prepared solution was mixed with 1.91 g ofdiethyl azodicarboxylate and stirred at room temperature for 1 hour.After distilling off the solvent, the resulting residue was purified bysubjecting it to silica gel column chromatography, thereby obtaining 1.2g of4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]-2-hydroxybenzaldehydein a viscous and oily form.

¹ H-NMR (CDCl₃) δ:1.47 (9H, s), 2.00-2.36 (2H, m), 3.30-3.75 (4H, m),4.94 (1H, quint), 6.38 (1H, d, J=2.1Hz), 6.52 (1H, dd, J=8.0 and 2.1Hz),7.44 (1H, d, J=8.0Hz), 9.72 (1H, s), 11.45 (1H, s)

b) 1.27 g of4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]-2-hydroxybenzaldehydeobtained in the above step a) and 0.884 g of ethyl bromoacetate weredissolved in 100 ml of acetone. The thus prepared solution was mixedwith 1.12 g of anhydrous potassium carbonate, and the mixture wasrefluxed for 1.5 hours. After cooling, insoluble materials were removedby filtration, and the solvent was distilled off. Thereafter, theresulting residue was purified by subjecting it to silica gel columnchromatography to obtain 1.44 g of ethyl[2-formyl-5-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]oxyacetatein a viscous and oily form.

¹ H-NMR (CDCl₃) δ:1.30 (3H, t, J=7.0Hz), 1.47 (9H, s), 2.00-2.28 (2H,m), 3.36-3.70 (4H, m), 4.28 (2H, q), 4.71 (2H, s), 4.94 (1H, quint),6.31 (1H, d, J=2.2Hz), 6.53 (1H, dd, J=8.8 and 2.2Hz), 7.84 (1H, s),10.39 (1H, s)

c) 1.44 g of ethyl[2-formyl-5-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]oxyacetateobtained in the above step b) and 1.71 g of(5-cyano-2-benzofuranyl)methyltriphenylphosphonium chloride weredissolved in a solvent mixture of 10 ml of tetrahydrofuran and 10 ml ofmethanol. The thus prepared solution was mixed with 0.664 g of1,8-diazabicyclo[5.4.0]-7-undecene, and the mixture was stirred for 1hour. After distilling off the solvent, the resulting residue waspurified by subjecting it to silica gel column chromatography using abenzene/ethyl acetate mixture as an eluant, thereby obtaining 1.8 g ofan olefinic compound as a mixture of E and Z forms. 1.8 g of the thusobtained olefinic compound was dissolved in a solvent mixture of 40 mlof tetrahydrofuran and 40 ml of ethanol, and the resulting solution wassubjected to catalytic hydrogenation under normal pressure in thepresence of 0.22 g of palladium oxide.barium sulfate.1H₂ O. Afterremoving the catalyst by filtration, the resulting residue was purifiedby subjecting it to silica gel column chromatography using abenzene/ethyl acetate mixture as an eluant. In this way, 1.6 g of thetitle compound was obtained in a viscous oily form. (Ester interchangewas effected during the reaction.)

¹ H-NMR (CDCl₃) δ:1.46 (9H, s), 1.96-2.08 (2H, m), 3.08 (4H, s),3.36-3.68 (4H, m), 3.80 (3H, m), 4.62 (2H, s), 4.80 (1H, br), 6.30 (1H,s), 6.36 (1H, d, J=8.0Hz), 6.43 (1H,s), 7.02 (1H, d, J=8.0Hz), 7.46 (2H,s), 7.77 (1H,s)

INVENTIVE EXAMPLE 1 ethyl3-(5-amidino-2-benzofuranyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride

1.96 g of ethyl2-[4-[((3S)-1-tert-butoxy-carbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-cyano-2-benzofuranyl)propionatewas dissolved in 150 ml of ethanol. With ice cooling and stirring,hydrogen chloride was bubbled into the thus prepared solution to asaturation level, followed by allowing it to stand for 18 hours. Theresulting reaction solution was concentrated to dryness under a reducedpressure, the thus obtained residue was dissolved in 300 ml of anethanol solution containing 15% (w/v) of ammonia and the solution wasallowed to stand still for 18 hours. After distilling off the solvent,the resulting residue was subjected to column chromatography using acolumn packed with a highly porous polymer type synthetic adsorbent(styrene-divinylbenzene polymer: Diaion HP-20) and using awater/acetonitrile mixture as an elution solvent. Fractions of interestthus pooled were subjected to reversed phase high performance liquidchromatography using a column packed with octadecyl-bonded silica geland using a water/acetonitrile mixture as an elution solvent.Thereafter, the thus eluted fractions of interest were pooled, mixedwith dilute hydrochloric acid, and then concentrated to dryness. In thisway, 610 mg of the title compound was obtained in a solid form.

¹ H-NMR (DMSO-d₆) δ:1.08 (3H, t, J=7.0), 1.90-2.30 (2H, m), 3.00-3.80(6H, m), 3.80-4.30 (3H, m), 5.08 (1H, br), 6.73 (1H, s), 6.93 (2H, d,J=8.3Hz), 7.33 (2H, d, J=8.3Hz), 7.73 (2H, s), 8.08 (1H, s), 9.25 (2H,br), 9.40 (2H, br), 9.50-10.00 (2H, br)

The following compounds of Inventive Examples 2 to 17 were prepared inaccordance with the procedure of Inventive Example 1.

INVENTIVE EXAMPLE 2 ethyl3-(5-amidino-2-benzofuranyl)-2-[4-[((2S,4S)-2-carbamoyl-4-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.10 (3H, t, J=7.0 Hz), 1.70-3.20 (2H, m), 3.00-4.50(8H, m), 5.00-5.30 (1H, br), 6.71 (1H, s), 6.87 (2H, d, J=8.3Hz), 7.30(2H, d, J=8.3Hz), 7.72 (2H, s), 8.10 (1H, s), 9.00-10.00 (6H)

INVENTIVE EXAMPLE 3 ethyl3-(5-amidino-2-benzofuranyl)-2-[4-[((2S,4S)-2-dimethylcarbamoyl-4-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.11 (3H, t, J=7.0Hz), 1.70-3.30 (2H, m), 2.91 (3H,s), 2.96 (3H, s), 3.00-4.20 (7H, m), 4.70 (1H, br), 5.10 (1H, br), 6.69(1H, s), 6.86 (2H, d, J=8.7Hz), 7.29 (2H, d, J=8.7Hz), 7.69 (2H, s),8.07 (1H, s), 8.80 (1H, br), 9.10 (2H, br), 9.34 (2H, br), 10.08 (1H,br)

INVENTIVE EXAMPLE 4 ethyl2-(5-amidino-2-benzofuranyl)-3-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.08 (3H, t, J=7.0Hz), 1.80-2.30 (2H, br), 2.70-3.70(6H, m), 4.08 (2H, q, J=7.0Hz), 4.35 (1H, t, J=7.9Hz), 5.08 (1H, br),6.84 (2H, d, J=8.3Hz), 6.96 (1H, s), 7.17 (2H, d, J=8.3Hz), 7.79 (2H,s), 8.12 (1H, s), 9.33 (2H, br), 9.51 (2H, br), 9.80 (2H, br)

INVENTIVE EXAMPLE 5 ethyl3-(5-amidino-2-benzofuranyl)-3-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.17 (3H, t, J=7Hz), 2.0-2.2 (2H, m), 3.0-3.8 (6H,m), 4.07 (2H, q), 4.5-4.7 (1H, m), 5.13 (1H, m), 6.94 (1H, s), 6.94 (2H,d, J=9Hz), 7.32 (2H, d, J=9Hz), 7.73 (2H, s), 8.13 (1H, s), 9.21 (2H,br), 9.40 (2H, br), 9.4-10.0 (2H, br)

INVENTIVE EXAMPLE 6 ethyl3-(5-amidino-3-benzofuranyl)-2-[4-[((3S)-3-pyrrodinyl)oxy]phenyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.06 (3H, t), 2.10 (2H, br), 3.0-3.7 (7H), 4.05 (2H,q), 5.09 (1H, br), 6.95 (2H, d), 7.28 (2H, d), 7.77 (3H), 8.21 (1H, s),9.2-9.8 (6H)

INVENTIVE EXAMPLE 7 ethyl2-[2-(5-amidino-2-benzofuranyl)ethyl]-5-[((3S)-3-pyrrolidinyl)oxy]benzoatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.29 (3H, t, J=7.0Hz), 2.00-2.35 (2H, m), 2.90-3.60(8H, m), 4.18 (2H, q, J=7.0Hz), 5.20 (1H, br), 6.75 (1H, s), 7.20 (1H,dd, J=7.9 and 2.8Hz), 7.39 (1H, d, J=7.9Hz), 7.41 (1H, d, J=2.8Hz), 7.74(2H, s), 8.09 (1H, s), 9.23 (2H, br), 9.40 (2H, br), 9.50-10.20 (2H, br)

INVENTIVE EXAMPLE 8 ethyl[2-[2-(5-amidino-2-benzofuranyl)ethyl]-5-[((3S)-3-pyrrolidinyl)oxy]phenyl]acetatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.18 (3H, t, J=7.0Hz), 2.0-2.30 (2H, m), 3.02 (4H,s), 3.00-4.00 (6H, m), 4.90 (2H, q, J=7.0Hz), 5.12 (1H, br), 6.80-7.00(3H, m), 7.24 (1H, d, J=8.64Hz), 7.76 (2H, s), 8.12 (1H, s), 9.29 (2H,br), 9.45 (2H, br), 9.40-10.10 (2H, br)

INVENTIVE EXAMPLE 9 ethyl5-amidino-2-[2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]ethyl]-3-benzofurancarboxylatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.36 (3H, t), 2.08 (2H, m), 4.30 (2H), 5.00 (1H,br), 6.90 (2H, d), 7.10 (2H, d), 7.83 (2H), 8.34 (1H, s), 9.27 (2H, br),9.51 (4H, br)

INVENTIVE EXAMPLE 10 ethyl3-(5-amidinobenzo[b]thien-2-yl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.19 (3H, t, J=7.0Hz), 2.00-2.40 (2H, m), 3.00-4.20(9H, m), 5.08 (1H, br), 6.92 (2H, d, J=8.75Hz), 7.27 (1H, s), 7.31 (2H,d, J=8.75Hz), 7.68 (1H, dd, J=8.3 and 1.5Hz), 8.10 (1H, d, J=8.3Hz),8.27 (1H, d, J=1.5Hz), 9.19 (2H, br), 9.42 (2H, br), 9.10-10.00 (2H, br)

INVENTIVE EXAMPLE 11 ethyl3-(6-amidinobenzo[b]thien-2-yl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.05 (3H, t), 2.10 (2H, m), 3.30 (7H, m), 4.0 (2H,q), 5.05 (1H, br), 6.90 (2H, d), 7.22 (3H, m), 7.60-7.90 (2H, m), 8.38(1H, s), 9.10 (2H, br), 9.35 (2H, br), 9.40 (2H, br)

INVENTIVE EXAMPLE 12 ethyl3-(6-amidino-1-ethyl-2-indolyl)-2-[4-[(4-piperidinyl)oxy]phenyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.05 (3H, t), 1.28 (3H), 1.83 (2H, br), 2.08 (2H,br), 2.90-3.20 (5H, br), 3.90-4.40 (3H, m), 4.62 (1H, br), 6.34 (1H, s),6.97(2H, d), 7.34 (2H), 7.47 (1H, d), 7.58 (1H, d), 8.13 (1H, s),8.90-9.40 (6H, br)

INVENTIVE EXAMPLE 13 ethyl3-(6-amidino-1-ethyl-2-indolyl)-2-[4-[((3R)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.08 (3H, t, J=7.0Hz), 1.31 (3H, t, J =7.0Hz),2.10-2.30 (2H, br), 3.17.(1H, dd), 3.20-3.40 (2H, m), 3.90-4.40 (5H, m),5.14 (1H, br), 6.37 (1H, s), 6.97 (2H, d, J=8.8Hz), 7.38 (2H, d,J=8.8Hz), 7.49 (1H, d, J=8.3Hz), 7.62 (1H, d, J=8.3Hz), 8.14 (1H, s),8.99 (2H, br), 9.32 (2H, br), 9.50-9.70 (2H, br)

INVENTIVE EXAMPLE 14 ethyl3-(6-amidino-1-methyl-2-indolyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride (ester interchange due to the use of ethanol solvent)

(solid)

¹ H-NMR (DMSO-d₆) δ:1.05 (3H, t), 1.95-2.30 (2H, m), 3.76 (3H, s), 4.02(2H, q), 4.00-4.30 (1H, m), 5.00-5.20 (1H, m), 6.38 (1H, s), 7.00 (2H,d), 7.40 (2H, d), 7.50-7.70 (2H, m), 8.25 (1H, s), 9.30-10.10(6H)

INVENTIVE EXAMPLE 15 ethyl3-(6-amidino-2-naphthyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.06 (3H, t, J=7.0Hz), 2.00-2.20 (2H, m), 3.00-4.00(7H, m), 3.99 (2H, q, J=7.0Hz), 5.11 (1H, m), 6.92 (2H, d, J=9.0Hz),7.31 (2H, d, J=9.0Hz), 7.55 (1H, d, J=8.0Hz), 7.80-8.10 (4H, m), 8.51(1H, s), 9.40 (2H, br), 9.58 (2H, br), 9.50-10.00 (2H, br)

INVENTIVE EXAMPLE 16 ethyl3-(7-amidino-2-naphthyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.01 (3H, t, J=7.0Hz), 2.00-2.20 (2H, m), 3.10-3.80(7H, m), 3.98 (2H, q, J=7.0Hz), 5.10 (1H, m), 6.93 (2H, d, J=9.0Hz),7.32 (2H, d, J=9.0Hz), 7.50-8.10 (5H, m), 8.44 (1H, m), 9.41 (2H, br),9.59 (2H, br), 9.30-10.00 (2H, br)

INVENTIVE EXAMPLE 17 ethyl3-(7-amidino-2-naphthyl)-2-[4-[(4-piperidinyl)methoxy]phenyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ: 1.01 (3H, t, J=7.1Hz), 1.45-1.55 (2H, m), 1.85-1.95(2H, m), 2.80-2.95 (2H, m), 3.15-3.50 (5H, m), 3.81 (2H, d), 3.95-4.05(2H, m), 4.05-4.15 (1H, m), 6.87 (2H, d, J=8.3Hz), 7.27 (2H, d,J=8.3Hz), 7.58-7.63 (1H, m), 7.75-7.80 (1H, m), 7.84 (1H, s), 7.95 (1H,d, J=8.8Hz), 8.07 (1H, d, J=8.8Hz), 8.40 (1H, s), 9.29 (2H), 9.53 (2H)

INVENTIVE EXAMPLE 183-(7-amidino-2-naphthyl)-2-[4-[(4-piperidinyl)methoxy]phenyl]propionicacid hydrochloride monohydrate

1.51 g of ethyl3-(7-amidino-2-naphthyl)-2-[4-[(4-piperidinyl)methoxy]phenyl]propionatedihydrochloride was dissolved in 50 ml of concentrated hydrochloricacid, and the solution was allowed to stand still in a sealed containerat room temperature for 62 hours. After drying the resulting reactionsolution under a reduced pressure, the thus obtained residue waspurified by applying it to a column packed with a highly porous polymertype synthetic adsorbent (styrene-divinylbenzene polymer: HP-20).Thereafter, the thus eluted fractions of interest were pooled, and mixedwith a small amount of ethanol, and then the crystals thus precipitatedwere collected by filtration. In this way, 0.79 g of the title compoundwas obtained in the form of crystals.

mp: 285°-287° C. (decomp.)

(Since solubility of the thus formed product to any solvent was verylow, the compound was made into dihydrochloride with hydrochloric acidand then dried prior to the NMR measurement.)

¹ H-NMR (DMSO-d₆) δ: 1.45-1.60 (2H, m), 1.85-1.95 (2H, m), 1.95-2.05(1H, br), 2.8-2.9 (2H, m), 3.1-3.2 (1H, m), 3.2-3.3 (2H, m), 3.4-3.5(1H, m), 3.80 (2H, d, J=6.4Hz), 3.9-4.0 (1H, m), 6.87 (2H, d, J=8.8Hz),7.27 (2H, d, J=8.8Hz), 7.60 (1H, d, J=8.l8Hz), 7.75-7.80 (1H, m), 7.83(1H, s), 7.94 (1H, d, J=8.8Hz), 8.07 (1H, d, J=8.3Hz) 8.40 (1H, s),8.8-8.9 (1H, br), 9.33 (2H), 9.54 (2H)

INVENTIVE EXAMPLE 193-(5-amidino-2-benzofuranyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

3.2 g of ethyl3-(5-amidino-2-benzofuranyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride was dissolved in 80 ml of 2N hydrochloric acid, and thesolution was refluxed under heating for 30 minutes. After cooling anddistilling off the solvent, the resulting residue was purified bysubjecting it to column chromatography using a column packed with ahighly porous polymer type synthetic adsorbent (styrene-divinylbenzenepolymer: Diaion HP-20) and using 5-10% acetonitrile as an elutionsolvent. Thereafter, the thus eluted fractions of interest were pooled,adjusted to pH 2-3 with dilute hydrochloric acid, and then concentratedto dryness. In this way, 1.25 g of the title compound was obtained in asolid form.

¹ H-NMR (DMSO-d₆) δ: 2.00-2.30 (2H, m), 3.00-3.80 (6H, m), 4.10 (1H, t,J=7.2Hz), 5.10 (1H, br), 6.74 (1H, s), 6.94 (2H, d, J=8.3Hz), 7.40 (2H,d, J=8.3Hz), 7.74 (2H, s), 8.09 (1H, s), 9.22 (2H, br), 9.40 (2H, br),9.10-10.00 (2H, br)

The following compounds of Inventive Examples 20 to 26 were prepared inaccordance with the procedure of Inventive Example 19.

INVENTIVE EXAMPLE 203-(5-amidino-2-benzofuranyl)-2-[4-[((2S,4S)-2-dimethylcarbamoyl-4-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ: 1.80-3.00 (2H, m), 2.89 (3H, s), 2.95 (3H, s),3.00-3.70 (4H, m), 4.09 (1H, t, J=7.9Hz), 4.70 (1H, br), 5.12 (1H, br),6.71 (1H, s), 6.86 (2H, d, J=8.3Hz), 7.30 (2H, d, J=8.3Hz), 7.73 (2H,s), 8.10 (1H, s), 8.76 (1H, br), 9.30 (2H, br), 9.46 (2H, br), 10.80(1H, br)

INVENTIVE EXAMPLE 212-(5-amidino-2-benzofuranyl)-3-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.90-2.30 (2H, m), 2.90-3.70 (6H, m), 4.26 (1H, t,J=7.9Hz), 5.06 (1H, br), 6.83 (2H, d, J=8.3Hz), 6.93 (1H, s), 7.17 (2H,d, J=8.3Hz), 7.78 (2H, s), 8.14 (1H, s), 9.30 (2H, br), 9.47 (2H, br),9.80 (2H, br)

INVENTIVE EXAMPLE 223-(5-amidino-2-benzofuranyl)-3-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ: 2.0-2.2 (2H, m), 3.0-4.0 (6H, m), 4.6 (1H, m), 5.10(1H, m), 6.92 (1H, s), 6.92 (2H, d, J=9.0Hz), 7.32 (2H, d, J=9.0Hz),7.73 (2H, s), 8.16 (1H, s), 9.30 (2H, br), 9.46 (2H, br), 9.6-10.0 (2H,br)

INVENTIVE EXAMPLE 232-[2-(5-amidino-2-benzofuranyl)ethyl]-5-[((3S)-3-pyrrolidinyl)oxy]benzoicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.90-2.30 (2H, m), 2.90-3.70 (8H, m), 4.96 (1H, br),6.75 (1H, s), 7.08 (1H, dd, =7.9 and 2.8Hz), 7.28 (1H, d, J=7.9Hz), 7.41(1H, d, =2.8Hz), 7.75 (2H, s), 8.09 (1H, s), 9.25 (2H, br), 9.42 (2H,br), 9.50-10.00 (2H, br)

INVENTIVE EXAMPLE 24[2-[2-(5-amidino-2-benzofuranyl)ethyl]-5-[((3S)-3-pyrrolidinyl)oxy]phenyl]aceticacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.95-2.30 (2H, m), 3.03 (4H, s), 5.04 (1H, br),6.68-6.90 (3H, m), 7.14 (1H, d, J=8.3Hz), 7.74 (2H, s), 8.10 (1H, s),9.38 (2H, br), 9.66 (2H, br), 9.00-10.00 (2H, br)

INVENTIVE EXAMPLE 253-(5-amidinobenzo[b]thien-2-yl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.90-2.40 (2H, m), 3.00-4.10 (7H, m), 5.14 (1H, br),6.93 (2H, d, J.=8.2Hz), 7.28 (1H, s), 7.33 (2H, d, J=8.2Hz), 7.70 (1H,d, J=8.8Hz), 8.09 (1H, d, J=8.8Hz), 8.26 (1H, s), 9.24 (2H, br), 9.47(2H, br), 9.00-10.20 (2H, br)

INVENTIVE EXAMPLE 263-(7-amidino-2-naphthyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ: 2.00-2.20 (2H, m), 3.00-3.70 (6H, m), 4.01 (1H, m),5.11 (1H, m), 6.92 (2H, d, J=9.0Hz), 7.33 (2H, d, J=9.0Hz), 7.50-8.20(5H, m), 8.43 (1H, s), 9.00-10.00 (2H, br)

INVENTIVE EXAMPLE 27 ethyl(+)-3-(7-amidino-2-naphthyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride

123.1 g of ethyl(+)-2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-cyano-2-naphthyl)propionatewas dissolved in a solvent mixture of 480 ml of dichloromethane and 1286ml of ethanol. With stirring at -10° C., hydrogen chloride was bubbledinto the thus prepared solution to a saturation level, and the resultingsolution was allowed to stand still for 26 hours at a temperature of -8°to -5° C. Thereafter, the resulting reaction solution was concentratedunder a reduced pressure at a temperature of 10° C. or below to obtain154 g of an oily material. The thus obtained oily material was dissolvedin 1480 ml of ethanol and, while keeping the inner temperature at -10°C. or below, ammonia gas was introduced until its concentration became21% (w/w) or more. After maintaining at a temperature of -8° to -5° C.for 107 hours, the resulting reaction solution was concentrated under areduced pressure at a temperature of 10° C. or below to distill off thesolvent, and the thus obtained residue was dissolved in 200 ml of water.After adjusting to pH 3-5 with dilute hydrochloric acid, the resultingsolution was purified by subjecting it to column chromatography using acolumn packed with a highly porous polymer type synthetic adsorbent(styrene-divinylbenzene polymer: Diaion HP-20) and using awater/acetonitrile mixture as an elution solvent. Thereafter, thuseluted fractions of interest were pooled, mixed with a small amount ofdilute hydrochloric acid, and then concentrated to dryness. In this way,107 g of the title compound was obtained in the form of a colorlesssolid.

¹ H-NMR (DMSO-d₆) δ:1.01 (3H, t, J=7.2Hz), 2.00-2.30 (2H, m), 3.1-3.6(6H, m), 3.90-4.05 (2H, m), 4.05-4.15 (1H, m), 5.10 (1H, br), 6.93 (2H,d, J=8.8Hz), 7.32 (2H), 7.60 (1H, d, J=8.3Hz), 7.78 (1H, d, J=8.3Hz),7.85 (1H, s), 7.96 (1H, d, J=8.3Hz), 8.08 (1H, d, J=8.3Hz), 8.41 (1H,s), 9.20-9.30 (2H, br), 9.40-9.70 (4H, br)

The following compounds of Inventive Examples 28 to 32 were prepared inaccordance with the procedure of Inventive Example 27.

INVENTIVE EXAMPLE 28 ethyl(-)-3-(7-amidino-2-naphthyl)-2-[4-[((3S)-3-pyrolidinyl)oxy]phenyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.02 (3H, t, J=7.2Hz), 2.00-2.30 (2H, m), 3.1-3.6(6H, m), 3.90-4.05 (2H, m), 4.05-4.15 (1H, m) 5.10 (1H, br), 6.94 (2H,d, J=8.8Hz), 7.32 (2H), 7.60 (1H, d, J=8.3Hz), 7.78 (1H, d, J=8.3Hz),7.86 (1H, s), 7.96 (1H, d, J=8.3Hz), 8.08 (1H, d, J=8.3Hz), 8.42 (1H,s), 9.20-9.30 (2H, br), 9.40-9.70 (4H, br)

INVENTIVE EXAMPLE 29 ethyl(+)-3-(5-amidinobenzo[b]thien-2-yl)-2-[4-[((2S)-2-pyrrolidinyl)methoxy]phenyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.10 (3H, t, J=7.3Hz), 1.73 (1H, dq, J=12.3 and8.3Hz), 1.84-2.05 (2H, m), 2.06-2.16 (1H, m), 3.12-3.27 (2H, br), 3.39(1H, dd, J=15.0 and 7.8Hz), 3.64 (1H, dd, J=15.0 and 7.8Hz), 3.80-3.93(1H, br), 4.00-4.24 (5H, m), 6.93 (2H, d, J=8.3Hz), 7.30 (1H, s), 7.31(2H, d), 7.67 (1H, d, J=8.3Hz), 8.11 (1H, d, J=8.3Hz), 8.23 (1H, s),9.12-9.30 (3H), 9.45 (2H, s), 9.43 (2H, s), 9.74-9.94 (1H, br)

INVENTIVE EXAMPLE 30 ethyl(-)-3-(5-amidinobenzo[b]thien-2-yl)-2-[4-[((2S)-2-pyrrolidinyl)methoxy]phenyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.09 (3H, t, J=7.3Hz), 1.72 (1H, dq, J=12.1 and8.3Hz), 1.84-2.03 (2H, m), 2.06-2.16 (1H, m), 3.12-3.27 (2H, br), 3.39(1H, dd, J=15.0 and 7.8Hz), 3.64 (1H, dd, J=15.0 and 7.8Hz), 3.80-3.93(1H, br), 4.00-4.24 (5H, m), 6.93 (2H, d, J=8.8Hz, 2 x ArH), 7.30 (1H,s), 7.31 (2H, d, J=8.8Hz), 7.67 (1H, dd, J=8.3 and 1.5Hz), 8.11 (1H, d,J=8.3Hz), 8.23 (1H, d, J=1.5Hz), 9.10-9.25 (1H, br), 9.21 (2H, s), 9.43(2H, s), 9.74-9.84 (1H, br)

INVENTIVE EXAMPLE 31 ethyl(+)-3-(7-amidino-2-naphthyl)-2-[4-[(4-piperidinyl)oxy]phenyl]propionatedihydrochloride

solid

¹ H-NMR (DMSO-d₆) 1.01 (3H, t, J=7.1Hz), 1.75-1.85 (2H, m), 2.05-2.15(2H, m), 3.0-3.1 (2H, m), 3.1-3.2 (3H, m), 3.9-4.0 (2H, m), 4.0-4.1 (1H,br), 4.1-4.2 (1H, m), 4.61 (1H, br), 6.95 (2H, d, J=8.8Hz), 7.29 (2H, d,J=8.8Hz), 7.61 (1H, d, J=8.3Hz), 7.78 (1H, d, J=8.3Hz), 7.84 (1H, s),7.95 (1H, d, J=8.3Hz), 8.07 (1H, d, J=8.8Hz), 8.38 (1H, s), 8.9-9.1 (2H,br), 9.20 (2H, br), 9.49 (2H, br)

INVENTIVE EXAMPLE 32 ethyl(-)-3-(7-amidino-2-naphthyl)-2-[4-[(4-piperidinyl)oxy]phenyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.01 (3H, t, J=7.1Hz), 1.75-1.80 (2H, m), 2.05-2.15(2H, m), 3.0-3.1 (2H, m), 3.1-3.3 (3H, m), 3.50-3.60 (1H, m), 3.65-3.75(2H, M), 3.9-4.0 (2H, m), 4.0-4.1 (1H, br), 4.1-4.2 (1H, m), 4.61 (1H,br), 6.95 (2H, d, J=8.3Hz), 7.29 (2H, d, J=8.3Hz), 7.61 (1H, d,J=8.8Hz), 7.78 (1H, d, J=8.3Hz), 7.84 (1H, s), 7.95 (1H, d, J=8.3Hz),8.07(1H, d, J=8.8Hz), 8.39 (1H, s), 8.9-9.1 (2H, br), 9.23 (2H, br),9.50 (2H, br)

INVENTIVE EXAMPLE 33 ethyl(+)-2-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionatedihydrochloride

In 1,000 ml of ethanol was dissolved 105.3 g of ethyl(+)-3-(7-amidino-2-naphthyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride. With stirring at room temperature, the thus preparedsolution was mixed with 51.5 g of ethyl acetimidate hydrochloride. Withice cooling and stirring, 89 ml of triethylamine was added dropwise tothe above solution while keeping the inner temperature at 3° to 5° C.,and the stirring was continued for 2.5 hours while keeping thetemperature at 5° C. or below. After distilling off the solvent under areduced pressure at a low temperature, the resulting reaction solutionwas adjusted to pH 4-5 with dilute hydrochloric acid, followed byfurther distillation under a reduced pressure to remove the solvent. Theresulting residue was purified by subjecting it to column chromatographyusing a column packed with a highly porous polymer type syntheticadsorbent (styrene-divinylbenzene polymer: Diaion HP-20) and using awater/acetonitrile mixture as an elution solvent. Thereafter, thuseluted fractions of interest were pooled, mixed with a small amount ofdilute hydrochloric acid, and then concentrated to dryness. In this way,110.1 g of the title compound was obtained in the form of a colorlesssolid.

¹ H-NMR (DMSO-d₆) δ: 1.02 (3H, m), 2.10-2.35 (2H, m), 2.26 (1.5H, s),2.31 (1.5H, s), 3.19 (1H, m), 3.40-3.85 (5H, m), 3.90-4.05 (2H, m),4.05-4.15 (1H, m), 5.13 (0.5H, br), 5.20 (0.5H, br), 6.90-6.97 (2H, m),7.32 (2H, m), 7.61 (1H, d, J=8.3Hz), 7.80 (1H, dd, J=8.3 and 1.5Hz),7.85 (1H, s), 7.96 (1H, d, J=8.3Hz), 8.08 (1H, d, J=8.3Hz), 8.43 (1H,s), 8.52 (0.5H, br), 8.61 (0.5H, br), 9.28-9.40 (3H, br), 9.50-9.60 (2H,br)

The following compounds of Inventive Examples 34 to 38 were prepared inaccordance with the procedure of Inventive Example 33.

INVENTIVE EXAMPLE 34 ethyl(-)-2-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-[7-amidino-2-naphthyl]propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.02 (3H, m), 2.10-2.35 (2H, m), 2.26 (1.5H, s),2.30 (1.5H, s), 3.19 (1H, m), 3.40-3.85 (5H, m), 3.90-4.05 (2H, m),4.05-4.15 (1H, m), 5.13 (0.5H, br), 5.20 (0.5H, br), 6.90-6.97 (2H, m),7.32 (2H, m), 7.61 (1H, d, J=8.3Hz), 7.80 (1H, dd, J=8.3 and 1.5Hz),7.84 (1H, s), 7.96 (1H, d, J=8.3Hz), 8.08 (1H, d, J=8.3Hz), 8.42 (1H,s), 8.52 (0.5H, br), 8.61 (0.5H, br), 9.28-9.40 (3H, br), 9.50-9.60 (2H,br)

INVENTIVE EXAMPLE 35 ethyl(+)-2-[4-[((2S)-1-acetimidoyl-3-pyrrolidinyl)methoxy]phenyl]-3-(5-amidinobenzo[b]thien-2-yl)propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.09 (3H, t, J=7.3Hz), 1.95-2.60 (4H, m), 2.26 (1H,s), 2.47 (2H, s), 3.30-3.70 (4H, m), 3.90-4.10 (5H, m), 4.40-4.60 (1H,m), 6.85-6.95 (2H, m), 7.28-7.33 (3H, m), 7.67 (1H, d, J=8.3Hz), 8.11(1H, d, J=8.3Hz), 8.23 (1H, s), 8.54 (2/3H, s), 8.69 (1/3H, s), 9.23(2H, s), 9.35-9.50 (3H, m)

INVENTIVE EXAMPLE 36 ethyl(-)-2-[4-[((2S)-1-acetimidoyl-3-pyrrolidinyl)methoxy]phenyl]-3-(5-amidinobenzo[b]thien-2-yl)propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ: 1.09 (3H, t, J=7.3Hz), 1.95-2.60 (4H, m), 2.26 (1H,s), 2.47 (2H, s), 3.30-3.70 (4H, m), 3.90-4.10 (5H, m), 4.40-4.60 (1H,m), 6.85-6.95 (2H, m),° 7.28-7.33 (3H, m), 7.67 (1H, d, J=8.3Hz), 8.10(1H, d, J=8.3Hz), 8.23 (1H, s), 8.51 (2/3H, s), 8.66 (1/3H, s), 9.16(2H, s), 9.30-9.48 (3H, m)

INVENTIVE EXAMPLE 37(+)-2-[4-[(1-acetimidoyl-4-piperidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionatedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ: 1.01 (3H, t, J=6.9Hz), 1.65-1.80 (2H, m), 2.0-2.1(2H, m), 2.30 (3H, s), 3.1-3.2 (1H, m), 3.2-3.8 (5H, m), 3.9-4.0 (2H,m), 4.0-4.1 (1H, br), 4.67 (1H, br), 6.95 (2H, d, J=8.8Hz), 7.30 (2H, d,J=8.8Hz), 7.61 (1H, d, J=8.3Hz), 7.78-7.84 (1H, m), 7.84 (1H, s), 7.95(1H, d, J=8.3Hz), 8.08 (1H, d, J=8.3Hz), 8.40 (1H, s), 8.80-9.55 (6H)

INVENTIVE EXAMPLE 38 ethyl(-)-2-[4-[(1-acetimidoyl-4-piperidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionatedihydrochloride

(solid)

[α]_(D) =-67.69° (c=0.585, H₂ O)

INVENTIVE EXAMPLE 39(+)-2-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid dihydrochloride

While keeping the inner temperature at -5° C. or below, 110.1 g of ethyl(+)-2-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionatedihydrochloride was dissolved in 3,300 ml of concentrated hydrochloricacid, and the resulting solution was allowed to stand still for 232hours at 5° C. The resulting reaction solution was concentrated bydistilling off hydrochloric acid and water under a reduced pressure. Theresulting residue was purified by subjecting it to column chromatographyusing a column packed with a highly porous polymer type syntheticadsorbent (styrene-divinylbenzene polymer: Diaion HP-20) and using awater/ acetonitrile mixture as an elution solvent. Thereafter, the thuseluted fractions of interest were pooled, mixed with a small amount ofdilute hydrochloric acid, and then concentrated to dryness. In this way,103.6 g of the title compound was obtained in the form of a light yellowsolid.

¹ H-NMR (DMSO-d₆) δ:2.10-2.4 (2H, m), 2.28 (1.5H, s), 2.31 (1.5H, s),3.10-3.30 (1H, m), 3.40-4.10 (6H, m), 5.14 (0.5H, br), 5.20 (0.5H, br),6.90-7.00 (2H, m), 7.35-7.40 (2H, m), 7.60 (1H, d, J=8.3Hz), 7.80 (1H,d, J=8.3Hz), 7.84 (1H, s), 7.94 (1H, d, J=8.3Hz), 8.06 (1H, d, J=8.3Hz),8.42 (1H, s), 8.55 (0.5H, br), 8.65 (0.5H, br), 9.30-9.70 (5H)

HPLC: Column; a ligand exchange type column with D-penicillamine as theoptically active site (SUMICHIRAL OA-5000, 4.6 φ×150 mm, Sumika AnalysisCenter)

Solvent;2 mM copper sulfate aqueous solution:

acetonitrile=85:15 (v/v)

Flow rate; 1 ml/min

Column temperature; 60° C.

Retention time; 43.60 minutes

The following compounds of Inventive Examples 40 to 44 were prepared inaccordance with the procedure of Inventive Example 39.

INVENTIVE EXAMPLE 40(-)-2-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:2.05-2.4 (2H, m), 2.28 (1.5H, s), 2.31 (1.5H, s),3.10-3.30 (1H, m), 3.40-4.10 (6H, m), 5.13 (0.5H, br), 5.20 (0.5H, br),6.90-7.00 (2H, m), 7.35-7.40 (2H, m), 7.60 (1H, d, J=8.3Hz), 7.81 (1H,d, J=8.3Hz), 7.84 (1H, s), 7.94 (1H, d, J=8.3Hz), 8.06 (1H, d, J=8.3Hz),8.42 (1H, s), 8.55 (0.5H, br), 8.64 (0.5H, br), 9.30-9.70 (5H)

HPLC: Column; a ligand exchange type column with D-penicillamine as theoptically active site (SUMICHIRAL OA-5000, 4.6 φ×150 mm, Sumika AnalysisCenter)

Solvent; 2 mM copper sulfate aqueous solution:

acetonitrile=85:15 (v/v)

Flow rate: 1 ml/min

Column temperature; 60° C.

Retention time; 38.14 minutes

INVENTIVE EXAMPLE 41(+)-2-[4-[((2S)-1-acetimidoyl-2-pyrrolidinyl)methoxy]phenyl]-3-(5-amidinobenzo[b]thien-2-yl)propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.95-2.60 (4H, m), 2.25 (1H, s), 2.44 (2H, s),3.15-3.80 (5H, m), 4.40-4.60 (1H, m), 6.83-6.95 (2H, m), 7.26 (1H, s),7.32 (2H, d, J=8.8Hz), 7.61 (1H, d, J=8.5Hz), 8.04 (1H, d, J=8.5Hz),8.21 (1H, s), 8.40-10.90 (6H)

INVENTIVE EXAMPLE 42(-)-2-[4-[((2S)-1-acetimidoyl-2-pyrrolidinyl)methoxy]phenyl]-3-(5-amidinobenzo[b]thien-2-yl)propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.95-2.60 (4H, m), 2.25 (1H, S), 2.44 (2H, s),3.15-3.75 (4H, m), 3.82 (1H, t, J=7.5Hz), 4.40-4.60 (1H, m), 6.83-6.95(2H, m), 7.27 (1H, s), 7.31 (2H, d, J=8.8Hz), 7.64 (1H, d, J=8.8Hz),8.06 (1H, d, J=8.8Hz), 8.2 1 (1H, s), 8.40-10.40 (6H)

INVENTIVE EXAMPLE 43(+)-2-[4-[(1-acetimidoyl-4-piperidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.65-1.80 (2H, m), 1.95-2.05 (2H, m), 2.31 (3H, s),3.1-3.2 (1H, m), 3.3-3.9 (5H, m), 3.95-4.05 (1H, m), 4.66 (1H, br), 6.95(2H, d, J=8.8HZ), 7.29 (2H, d, J=8.3Hz), 7.60 (1H, d, J=8.8Hz), 7.81(1H, d, J=8.8Hz), 7.84 (1H, s), 7.95 (1H, d, J=8.8Hz), 8.07 (1H, d,J=8.8Hz), 8.43 (1H, s), 8.80-9.65 (6H)

INVENTIVE EXAMPLE 44(-)-2-[4-[(1-acetimidoyl-4-piperidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.65-1.80 (2H, m), 2.00-2.10 (2H, m), 2.30 (3H, s),3.1-3.2 (1H, m), 3.3-3.85 (5H, m), 3.95-4.05 (1H, m), 4.66 (1H, m), 6.95(2H, d, J=8.8Hz), 7.30 (2H, d, J=8.8Hz), 7.61 (1H, d, J=8.8Hz), 7.78(1H, d, J=8.8Hz), 7.85 (1H, s), 7.95 (1H, d, J=8.8Hz), 8.08 (1H, d,J=8.8Hz), 8.40 (1H, s), 8.60-9.65 (6H)

INVENTIVE EXAMPLE 45(+)-2-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid hydrochloride pentahydrate

102.6 g of(+)-2-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid dihydrochloride was dissolved in 1,000 ml of water. With stirring,the thus prepared solution was adjusted to pH 4.8 by gradually adding astrongly basic OH type ion exchange resin (Amberlite IRA-410).Thereafter, the resin was removed by filtration, and the resultingfiltrate was concentrated to dryness. The thus obtained residue (94.6 g)was dissolved in 142 ml of water, and the solution was mixed with 1,570ml of ethanol, and stirred at room temperature for 1 hour. Afterremoving thus formed crystals by filtration, the resulting mother liquorwas seeded with the crystals of interest, and stirred at 8° C. for 40hours. Thereafter, the thus precipitated crystals were collected bysuction filtration, washed with ethanol, and then air-dried for 6.5hours under normal pressure at a relative humidity of 60 to 70%. In thisway, 70.3 g of the title compound was obtained in the form of colorlessprism crystals.

[α]_(D) ²⁴ =+57.4° (C=1.000, H₂ O) (solubilization at 40° C., measuredafter 30 minutes of heating at this temperature)

¹ H-NMR (DMSO-d₆) δ:2.20-2.35 (2H, m), 2.29 (1.5H, s), 2.32 (1.5H, s),2.80-2.95 (1H, m), 3.30-4.00 (6H, m), 5.16 (0.5H, br), 5.22 (0.5H, br),6.90-7.00 (2H, m), 7.45-7.51 (2H, m), 7.57 (1H, d, J=8.3Hz), 7.66 (1H,d, J=8.0Hz), 7.93 (1H, d, J=8.3Hz), 7.97 (1H, d, J=8.3Hz), 8.11 (1H, s),8.68 (1H, br), 8.70-9.30 (br), 11.50-12.20 (br)

Elementary analysis

theoretical value (C₂₆ H₂₈ N₄ O₃.HCl.5H₂ O): C, 54.68; H, 6.88; N, 9.80;Cl, 6.21

analytical value: C, 54.77; H, 6.76; N, 9.68; Cl, 6.42

Based on the results of crystal X-ray analysis, the thus obtained titlecompound was judged to be(2S)-2-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid

INVENTIVE EXAMPLE 46 methyl(+)-3-(6-amidino-2-indolyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride

During ice cooling, hydrogen chloride was bubbled into a solvent mixtureof 10 ml of dichloromethane and 20 ml of methanol. To the thus saturatedsolution was added 10 ml of a dichloromethane solution containing 450 mgof (+)-((2S)-1-p-toluenesulfonyl-2-pyrrolidinyl)methyl2-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-cyano-2-indolyl)propionate.The thus prepared mixture was allowed to stand still for 72 hours at 5°C. After concentration to dryness under a reduced pressure at atemperature of 40° C. or below, the resulting residue was dissolved in20 ml of ethanol solution containing 14% (w/v) of ammonia and stirred atroom temperature for 24 hours. After distilling off the solvent, thethus obtained residue was subjected to reversed phase high performanceliquid chromatography using a column packed with octadecyl-bonded silicagel and using a water/acetonitrile mixture as an elution solvent.Thereafter, the thus eluted fractions of interest were pooled, mixedwith dilute hydrochloric acid, and then concentrated to dryness. In thisway, 95 mg of the title compound was obtained in a solid form.

¹ H-NMR (DMSO-d₆) δ:2.30 (2H, m), 3.30 (1H, dd), 3.50-3.60 (5H, m), 3.70(3H, s), 4.20 (1H, t), 5.20 (1H, m), 6.33 (1H, s), 6.96 (2H, d), 7.33(2H, d), 7.40 (1H, d), 7.64 (1H, d), 7.80 (1H, s), 9.30-9.80 (6H, m)

INVENTIVE EXAMPLE 47 methyl(-)-3-(6-amidino-2-indolyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride

This compound was prepared in accordance with the procedure of InventiveExample 46.

(solid)

¹ H-NMR (DMSO-d₆) δ:2.30 (2H, m), 3.30 (1H, dd), 3.50-3.60 (5H, m), 3.70(3H, s), 4.20 (1H, t), 5.20 (1H, m), 6.33 (1H, s), 6.96 (2H, d), 7.33(2H, d), 7.40 (1H, d), 7.64 (1H, d), 7.80 (1H, s), 9.30-9.80 (6H, m)

INVENTIVE EXAMPLE 48(+)-3-(6-amidino-2-indolyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

1.8 g of methyl(+)-3-(6-amidino-2-indolyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride was dissolved in. 60 ml of concentrated hydrochloricacid, and the solution was stirred at 5° C. for 7 days. Afterconcentrating the resulting reaction solution to dryness under a reducedpressure at a temperature of 50° C. or below, the thus obtained residuewas subjected to reversed phase high performance liquid chromatographyusing a column packed with octadecyl-bonded silica gel and using awater/acetonitrile mixture as an elution solvent. Thereafter, thuseluted fractions of interest were pooled, mixed with dilute hydrochloricacid, and then concentrated to dryness. In this way, 1.3 g of the titlecompound was obtained in a solid form.

IR (KBr): 3600-3300, 1730, 1680 cm⁻¹

INVENTIVE EXAMPLE 49(-)-3-(6-amidino-2-indolyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

This compound was prepared in accordance with the procedure of InventiveExample 48.

(solid)

IR (KBr): 3600-3300, 1730, 1680 cm ⁻¹

INVENTIVE EXAMPLE 503-(5-amidino-2-benzofuranyl)-2-[4-[((3S)-1-methyl-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

1.0 g of ethyl3-(5-cyano-2-benzofuranyl)-2-[4-[((3S)-1-methyl-3-pyrrolidinyl)oxy]phenyl]propionatewas dissolved in 70 ml of ethanol. During ice cooling and stirring,hydrogen chloride was bubbled into the thus prepared solution to asaturation level. The thus saturated solution was allowed to stand stillat 25° C. for 20 hours. After distilling off the solvent under a reducedpressure, the thus obtained residue was dissolved in 50 ml of ethanolcontaining 14% (w/v) of ammonia, and the resulting solution was allowedto stand still at 25° C. for 20 hours. Thereafter, the solvent wasremoved by distillation to obtain ethyl3-(5-amidino-2-benzofuranyl)2-[4-[((3S)-1-methyl-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride. The thus obtained ester compound was dissolved in 50 mlof 2N hydrochloric acid and refluxed under heating for 30 minutes. Afterdistilling off the solvent under a reduced pressure, the resultingresidue was subjected to column chromatography using a column packedwith a highly porous polymer type synthetic adsorbent(styrene-divinylbenzene polymer: Diaion HP-20) and using awater/acetonitrile mixture as an elution solvent. Fractions of interestthus pooled were subjected to reversed phase high performance liquidchromatography using a column packed with octadecyl-bonded silica geland using a water/acetonitrile mixture as an elution solvent.Thereafter, thus eluted fractions of interest were pooled, mixed withdilute hydrochloric acid, and then concentrated to dryness. In this way,200 mg of the title compound was obtained in a solid form.

¹ H-NMR (DMSO-d₆) δ:2.40-3.40 i(6H, m), 2.92 (3H, m), 5.10-5.40 (1H,br), 6.82 (1H, s), 7.01 (2H, d, J=8.4Hz), 7.43 (2H, d, J=8.4Hz), 7.82(2H, s), 8.17 (1H, s), 9.34 (2H, br), 9.53 (2H, br)

The following compounds of Inventive Examples 51 to 82 were prepared inaccordance with the procedure of Inventive Example 50.

INVENTIVE EXAMPLE 512-[4-[((3S)-1-acetyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-amidino-2-benzofuranyl)propionic acid hydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.90-2.38 (5H, m), 3.00-3.90 (6H, m), 4.06 (1H, t,J=7.2Hz), 4.88 (1H, br), 6.67 (1H, s), 6.87 (2H, d, J=8.3Hz), 7.29 (2H,d, J=8.3Hz), 7.70 (2H, s), 8.08 (1H, s), 9.20 (2H, br), 9.41 (2H, br)

INVENTIVE EXAMPLE 523-(5-amidino-2-benzofuranyl)-2-[4-[((3S)-1-dimethylcarbamoyl-3-pyrrolidinyl)oxy]phenyl]propionicacid hydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.84-2.20 (2H, m), 2.75 (6H, s), 3.00-3.90 (6H, m),4.09 (1H, t, J=7.2Hz), 4.87-5.10 (1H, br), 6.68 (1H, s), 6.87 (2H, d,J=8.75Hz), 7.29 (2H, d, J=8.75Hz), 7.70 (2H, s), 8.07 (1H, s), 9.23 (2H,br), 9.39 (2H, br)

INVENTIVE EXAMPLE 533-(5-amidino-2-benzofuranyl)-2-[4-[((2S)-2-pyrrolidinyl)methoxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.95 (4H, br), 6.71 (1H, s), 6.97 (2H, d), 7.27 (2H,d), 7.71 (2H, s), 8.06 (1H, s), 9.15-9.35 (5H), 9.7 (1H)

INVENTIVE EXAMPLE 543-(5-amidino-2-benzofuranyl)-2-[4-[(tetrahydro-3-furanyl)oxy]phenyl]propionicacid hydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.6-2.4 (2H, m), 3.0-3.9 (6H, m), 4.0 (1H, dd),4.8-5.1 (1H, m), 6.75 (1H, s), 6.9 (2H, d), 7.32 (2H), 7.77 (2H, s), 8.1(1H, s), 9.37 (4H, d)

INVENTIVE EXAMPLE 553-(5-amidino-3-methyl-2-benzofuranyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:2.06 (5H, m), 5.05 (1H, br), 6.94 (2H, d), 7.22 (2H,d), 7.70 (2H, s), 8.08 (1H, s), 9.10-9.50 (6H, m)

INVENTIVE EXAMPLE 563-(5-amidino-7-methoxy-2-benzofuranyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.90-2.40 (2H, m), 2.90-3.80 (6H, m), 4.03 (3H, s),5.00-5.20 (1H, br), 6.65 (1H, s), 6.91 (2H, d, J=8.3Hz), 7.31 (2H, d,J=8.3Hz), 7.32 (1H, s), 7.68 (1H, s), 9.16 (2H, br), 9.40 (2H, br),9.20-10.0 (2H, br)

INVENTIVE EXAMPLE 573-(5-amidino-3-benzofuranyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:2.1 (2H, br), 3.00-4.00 (7H, m), 5.08 (1H, br), 6.90(2H, d, J=8Hz), 7.30 (2H, d, J=8Hz), 7.77 (3H), 8.22 (1H, s), 9.0-10.00(6H)

INVENTIVE EXAMPLE 585-amidino-2-[2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]ethyl]-3-benzofurancarboxylicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ: 2.07 (2H, m), 3.00-3.50 (8H), 5.05 (1H, br), 6.85(2H, d, J=8.0Hz), 7.15 (2H, d, J=8.0Hz), 7.82 (2H, s), 8.35 (1H, s),9.30 (2H, br), 9.50 (4H, br)

INVENTIVE EXAMPLE 593-[2-[2-(5-amidinobenzo[b]thien-2-yl)ethyl]-4-ethoxy-5-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.30 (3H, t, J=7.0Hz), 2.00-3.90 (14H, m), 4.01 (2H,q, J=7.0Hz), 6.94 (1H, s), 6.96 (1H, s), 7.39 (1H, s), 7.79 (1H, d,J=9.0Hz), 8.20 (1H, d, J=9.0Hz), 8.37 (1H, s), 9.41 (2H, br), 9.59 (2H,br), 9.0-10.0 (2H, br)

INVENTIVE EXAMPLE 603-[2-[2-[5-amidinobenzo[b]thien-2-yl)ethyl]-5-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:2.00-4.80 (14H, m), 5.08 (1H, m), 6.77 (1H, d,J=8.5Hz), 6.82 (1H, s), 7.18 (1H, d, J=8.5Hz), 7.35 (1H, s), 7.72 (1H,d, J=8.7Hz), 8.16 (1H, d, J=8.7Hz), 8.29 (1H, s), 9.31 (2H, br), 9.51(2H, br), 9.3-9.8 (2H, br)

INVENTIVE EXAMPLE 614-(5-amidinobenzo[b]thien-2-yl)-3-[4-[((3S)-3-pryrrolidinyl)oxy]phenyl]butyricacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ: 2.00-2.30 (2H, m), 5.00-5.20 (1H, m), 6.85 (2H, d),7.20 (1H, S), 7.25 (2H, d), 7.65 (1H, dd), 8.05-8.25 (2H, m)

INVENTIVE EXAMPLE 623-(5-amidinobenzo[b]thien-2-yl)-2-[4-[[2-(ethoxycarbonylimino)hexahydropyrimidine-5-yl]oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.27 (3H, t, J=7.0Hz), 3.00-4.04 (br), 4.24 (2H, q,J=7.0Hz), 4.90-5.10 (1H, br), 6.99 (2H, d, J=8.3Hz), 7.34 (2H, d,J=8.3Hz), 7.39 (1H, s), 7.68 (1H, dd, J=9.0 and 1.8Hz), 8.10 (1H, d,J=9.0Hz), 8.24 (1H, d, J=1.8Hz), 8.98 (2H, br), 9.23 (2H, br), 9.44 (2H,br), 11.65 (1H, s)

INVENTIVE EXAMPLE 633-(5-amidinobenzo[b]thien-2-yl)-2-[4-[[2-(imino)hexahydropyyrimidine-5-yl]oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:3.20-4.20 (3H, m), 3.44 (4H), 4.80-5.00 (1H, br),6.98 (2H, d, J=8.31Hz), 7.17 (2H, s), 7.29 (1H, s), 7.34 (2H, d,J=8.31Hz), 7.70 (1H, dd, J=8.2 and 2.0Hz),8.06 (2H, s), 8.12 (1H, d,J=8.2Hz), 8.25 (1H, s), 9.46 (2H, br), 9.57 (2H, br)

INVENTIVE EXAMPLE 643-(5-amidinobenzo[b]thien-2-yl)-2-[4-[((2S)-2-pyrrolidinyl)methoxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.95 (4H, m), 3.00-4.20 (8H, m), 6.95 (2H, d,J=8.0Hz), 7.28 (3H), 7.70 (1H, d, J=8.0Hz), 8.06 (1H, d, J=8.0Hz), 8.23(1H, s), 9.20-9.50 (6H)

INVENTIVE EXAMPLE 653-(5-amidinobenzo[b]thien-2-yl)-2-[4-[(4-piperidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) 1.80-2.15 (4H, m), 3.00-3.25 (4H, m), 3.30-4.00 (3H,m), 4.60-4.70 (1H, m), 6.97 (2H, d, J=8.3Hz), 7.31 (3H, m), 7.69 (1H,dd), 8.13 (1H, d, J=8.8Hz), 8.26 (1H, s), 9.31 (2H, br), 9.50 (2H, br),9.00-10.00 (2H, br)

INVENTIVE EXAMPLE 663-(5-amidinobenzo[b]thien-2-yl)-2-[4-[(2-aminoethyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:3.00-4.40 (7H, m), 6.93 (2H, d, J=8.3Hz), 7.29 (1H,s), 7.32 (2H, d, J=8.3Hz), 7.67 (1H, dd, J=9.0 and 1.0Hz), 8.20 (1H, d,J=9.0Hz), 8.32 (1H, s), 8.10-8.60 (3H, br), 9.24 (2H, br), 9.46 (2H, br)

INVENTIVE EXAMPLE 673-(5-amidinobenzo[b]thien-2-yl)-2-[4-[2-(1-pyrrolin-2-yl)aminoethoxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.88-2.30 (2H, m), 2.60-3.00 (2H, m), 3.00-4.30 (9H,m), 6.90 (2H, d, J=8.3Hz), 7.30 (1H, s), 7.31 (2H, d, J=8.3Hz), 7.70(1H, dd, J=8.50 and 1.00Hz), 8.11 (1H, d, J=8.50Hz), 8.25 (1H, s), 9.28(2H, br), 9.48 (2H, br), 10.00 (1H, br),. 10.19 (1H, br)

INVENTIVE EXAMPLE 683-(5-amidino-2-indolyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:2.00-2.35 (2H, m), 4.00-4.30 (1H, m), 5.00-5.30 (1H,br), 6.37 (1H, s), 7.00 (2H, d), 7.40 (2H, d), 7.60 (2H, d), 8.10 (1H,s), 11.60 (1H, s)

INVENTIVE EXAMPLE 69

3-(6-amidino-2-indolyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:2.20 (2H, m), 3.40 (4H, m), 5.16 (1H, br), 6.36 (1H,s), 7.00 (2H, d), 7.27 (2H, d), 7.36-7.96 (3H, m), 9.20-9.50 (6H, m),11.80 (1H, s)

INVENTIVE EXAMPLE 703-(5-amidino-2-indolyl)-2-[4-[((3R)-tetrahydro-3-furanyl)oxy]phenyl]propionicacid hydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:5.00 (1H, br), 6.28 (1H, s), 6.82 (2H, d), 7.30 (2H,d), 7.58 (2H, s), 8.00 (1H, s), 9.10 (4H), 11.8 (1H, s)

INVENTIVE EXAMPLE 713-(5-amidino-2-indolyl)-2-[4-[((3S)-tetrahydro-3-furanyl)oxy]phenyl]propionicacid hydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:5.10 (1H, br), 6.27 (1H, s), 6.82 (2H, d), 7.29 (2H,d), 7.58 (2H, s), 8.00 (1H, s), 9.12 (4H), 11.8 (1H, s)

INVENTIVE EXAMPLE 72 3-(5-amidino-1-methyl-2-indoly)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.90-2.25 (2H, m), 3.73 (3H, s), 5.00-5.20 (1H, br),6.40 (1H, s), 6.95 (2H, d), 7.40 (2H, d), 7.62 (2H, s), 8.10 (1H, s),9.00-9.80 (6H, br)

INVENTIVE EXAMPLE 733-(6-amidino-1-ethyl-2-indolyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.30 (3H, t), 1.95-2.30 (2H, m), 5.10 (1H, m), 6.37(1H, s), 6.92 (2H, d), 7.30-7.70 (4H, m), 8.10 (1H, s), 9.30-9.90 (6H,br)

INVENTIVE EXAMPLE 743-(6-amidinol-1-ethyl-2-indolyl)-2-[4-[((3R)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.31 (3H, t), 2.00-4.50 (11H), 5.11 (1H, br), 6.38(1H, s), 6.96 (2H, d, =8.4Hz), 7.30-7.70 (4H, m), 8.17 (1H, s), 9.07(2H, br), 9.34 (2H, br), 9.30-10.00 (2H, br)

INVENTIVE EXAMPLE 753-[6-amidino-1-(2-chloroethyl)-2-indolyl]-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:2.00-5.00 (13H), 5.13 (1H, br), 6.42 (1H, s), 6.97(2H, d, J=8.4Hz), 7.40 (2H, d, J=8.4Hz), 7.50-7.70 (2H, m), 8.22 (1H,s), 9.13 (2H, br), 9.39 (2H, br), 9.50-10.00 (2H, br)

INVENTIVE EXAMPLE 763-(6-amidino-1,2,3,4-tetrahydro-2-naphthyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ: 1.30-4.00 (16H, m), 5.10 (1H, m), 6.94 (2H, d,J=9.0Hz), 7.20-7.70 (5H, m), 9.18 (2H, br), 9.34 (2H, br), 9.50-10.00(2H, br)

INVENTIVE EXAMPLE 773-(5-amidino-2-benzimidazolyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.98-2.28 (2H, br), 3.00-4.80 (7H, m), 5.00-5.20(1H, br), 6.93 (2H, d, J=9.0Hz), 7.34 (2H, d, J=9.0Hz), 7.91 (2H, s),8.28 (1H, s), 9.36 (2H, br), 9.61 (2H, br), 9.40-10.10 (2H, br)

INVENTIVE EXAMPLE 783-(7-amidino-2-naphthyl)-2-[4-[((3R)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ: 2.00-2.20 (2H, m), 3.00-4.20 (7H, m), 5.10 (1H,br), 6.92 (2H, d, J=9.0Hz), 7.33 (2H, d, J=9.0Hz), 7.50-8.20 (5H, m),8.43 (1H, s), 9.39 (2H, br), 9.60 (2H, br), 9.50-10.00 (2H, br)

INVENTIVE EXAMPLE 793-(7-amidino-2-naphthyl)-2-[4-[(4-piperidinyl)oxy]phenyl]propionic aciddihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ: 1.70 -2.20 (4 H, m ), 2.80 -4.10 (7H, m), 4.50-4.80(1H, m), 6.95 (2H, d, J=8.0Hz), 7.30 (2H, d, J =8.0Hz), 7.60-8.50 (6H,m), 9.35 (2H, br), 9.57 (2H, br), 9.10-9.80 (2H, br)

INVENTIVE EXAMPLE 803-(6-amidino-1-carboxyethyl-2-indolyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ: 1.98-2.30 (2H, m), 2.80-3.80 (6H, m), 3.90-4.25(1H, t), 5.00-5.50 (3H, br), 6.41 (1H, s), 7.00 (2H, d), 7.42 (2H, d),7.60-7.90 (2H, m), 8.30 (1H, s), 9.10-10.00 (6H, br)

INVENTIVE EXAMPLE 816-amidino-2-[3-hydroxy-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propyl]-1-indoleaceticacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.95-2.20 (2H, m), 4.90-5.15 (3H, m), 6.20 (1H, s),6.90 (2H, d), 7.25 (2H, d), 7.57 (2H, m), 8.20 (1H, s), 9.20-9.90 (6H,br)

INVENTIVE EXAMPLE 826-amidino-2-[2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]ethyl]-1-indoleaceticacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.90-2.30 (2H, m), 3.10-3.50 (4H, 4.80-5.30 (3H,br), 6.42 (1H, s), 6.90 (2H, d), 7.25 (2H, d), 7.60 (3H, m), 8.25 (1H,s), 9.20-10.00 (6H, br)

INVENTIVE EXAMPLE 83 ethyl3-(5-amidinobenzo[b]thien-2-yl)-2-ethoxycarbonyl-2-[4-[((2R)-2-pyrrolidinyl)methoxy]phenyl]propionatedihydrochloride

4.34 g of ethyl 2-[4-[((2R)-1-tert-butoxy-carbonyl-2-pyrrolidinyl)methoxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)-2-ethoxycarbonylpropionatewas dissolved in 150 ml of ethanol. During ice cooling and stirring,hydrogen chloride was bubbled into the thus prepared solution to asaturation level. The thus saturated solution was allowed to stand stillat room temperature for 18 hours. After distilling off the solvent undera reduced pressure, the thus obtained residue was dissolved in 100 ml ofethanol solution containing 13% (w/v) of ammonia, and the resultingsolution was allowed to stand still for 24 hours. After distilling offthe solvent, the resulting residue was subjected to columnchromatography using a column packed with a highly porous polymer typesynthetic adsorbent (styrene-divinylbenzene polymer: Diaion HP-20) andusing a water/acetonitrile mixture as an elution solvent. Fractions ofinterest thus pooled were subjected to reversed phase high performanceliquid chromatography using a column packed with octadecyl-bonded silicagel and using a water/acetonitrile mixture as an elution solvent.Thereafter, the thus eluted fractions of interest were pooled, mixedwith dilute hydrochloric acid and then concentrated to dryness. In thisway, 1.0 g of the title compound was obtained in the form of lightyellow solid.

¹ H-NMR (DMSO-d₆) δ:1.15 (6H, t, J=7.0Hz), 2.0 (4H, br), 3.00-4.00 (3H),3.95 (2H), 4.2 (4H), 7.00 (2H, d), 7.16 (1H), 7.31 (2H, d), 7.70 (1H,dd), 8.10 (1H, d), 8.26 (1H, d), 9.20-9.60 (5H), 9.9 (1H)

INVENTIVE EXAMPLE 843-(5-amidinobenzo[b]thien-2-yl)-2-[4-[(2-imidazolin-2-yl)methoxy]phenyl]propionicacid dihydrochloride

1.6 g of ethyl3-(5-cyanobenzo[b]thien-2-yl)-2-ethoxycarbonyl-2-[4-[(2-imidazolin-2-yl)methoxy]phenyl]propionatewas dissolved in 100 ml of ethanol. During ice cooling and stirring,hydrogen chloride was bubbled into the thus prepared solution to asaturation level. The thus saturated solution was allowed to stand stillat 5° C. for 18 hours. After distilling off the solvent under a reducedpressure, the thus obtained residue was dissolved in 100 ml of ethanolsolution containing 13% (w/v) of ammonia, and the resulting solution wasallowed to stand still at room temperature for 24 hours. Thereafter, thesolvent was distilled off to obtain ethyl3-(5-amidinobenzo[b]thien-2-yl)-2-ethoxycarbonyl-2-[4-[(2-imidazolin-2-yl)methoxy]phenyl]propionatedihydrochloride. The thus obtained ester compound was dissolved in 50 mlof 5N hydrochloric acid, and the solution was refluxed under heating for1 hour. After distilling off the solvent, the resulting residue wassubjected to column chromatography using a column packed with a highlyporous polymer type synthetic adsorbent (styrene-divinylbenzene polymer:Diaion HP-20) and using a water/acetonitrile mixture as an elutionsolvent. Fractions of interest thus pooled were subjected to reversedphase high performance liquid chromatography using a column packed withoctadecyl-bonded silica gel and using a water/acetonitrile mixture as anelution solvent. Thereafter, the thus eluted fractions of interest werepooled, mixed with dilute hydrochloric acid, and then concentrated todryness. In this way, 200 mg of the title compound was obtained in theform of a light yellow solid.

¹ H-NMR (DMSO-d₆) δ:3.35 (1H, dd), 3.65 (1H, dd), 3.89 (4H, s), 3.99(1H, t), 5.07 (2H, s), 6.98 (2H, d), 7.32 (1H, s), 7.37 (2H, d), 7.66(1H, d), 8.12 (1H, d), 8.21 (1H, s), 9.10 (2H), 9.39 (2H), 10.38 (2H).

The following compounds of Inventive Examples 85 and 86 were prepared inaccordance with the procedure of Inventive Example 82.

INVENTIVE EXAMPLE 853-(5-amidinobenzo[b]thien-2-yl)-2-[4-[((3S)-3-pyrrolidinyl)thio]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.5-4.5 (10H, m), 6.95 (2H, d), 7.32 (1H, s), 7.40(2H, d), 7.71 d), 8.13 (1H, d), 8.28 (1H, s), 9.3 (2H, br), 9.5 (2H,br), 9.8 (2H, br)

INVENTIVE EXAMPLE 863-(5-amidino-2-benzothiazolyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:2.08 (2H, br), 3.00-4.25 (7H), 5.10 (1H, br), 6.95(2H, d),-7.34 (2H, d), 7.82 (1H, dd), 8.29 (1H, d), 8.41 (1H, d),9.20-9.60 (6H)

FAB MS (m/z): 411 (M⁺ +1)

INVENTIVE EXAMPLE 872-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-amidino-2-benzofuranyl)propionicacid dihydrochloride

1.1 g of3-(5-amidino-2-benzofuranyl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride was dissolved in 20 ml of water. During ice coolingand stirring, 1.4 g of ethyl acetimidate hydrochloride was addedgradually to the thus prepared solution while adjusting the pH of thesolution to 8.5 with 1N sodium hydroxide aqueous solution. The resultingmixture was stirred for 15 minutes with ice cooling, and then adjustedto pH 2.0 with dilute hydrochloric acid. After concentrating theresulting reaction solution to dryness, the thus obtained residue wassubjected to reversed phase high performance liquid chromatography usinga column packed with octadecyl-bonded silica gel and using awater/acetonitrile mixture as -an elution solvent. Thereafter, the thuseluted fractions of interest were pooled, mixed With dilute hydrochloricacid, and then concentrated to dryness. In this way, 780 mg of the titlecompound was obtained in a solid form.

¹ H-NMR (DMSO-d₆) δ: 1.90-2.40 (5H, m), 2.90-4.30 (7H, br), 4.96 (1H,br), 6.73 (1H, s), 6.93 (2H, d, J=8.8Hz), 7.33 (2H, d, J=8.8Hz), 7.73(2H, s), 8.10 (1H, s), 8.50-8.80 (1H, br), 9.33 (2H, br), 9.46 (3H, br)

The following compounds of Inventive Examples 88 to 91 were prepared inaccordance with the procedure of Inventive Example 87.

INVENTIVE EXAMPLE 882-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-amidinobenzo[b]thien-2-yl)propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:2.00-2.50 (5H, m), 3.10-4.20 (7H, m), 4.96 (1H, br),6.93 (2H, d, J=7.9Hz), 7.2 (1H, s), 7.34 (2H, d, J=7.9Hz), 7.73 (1H, d,J=8.3Hz), 8.10 (1H, d, J=8.3Hz), 8.30 (1H, s), 8.50-9.30 (1H, br), 9.37(2H, br), 9.54 (2H, br)

INVENTIVE EXAMPLE 892-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(5-amidino-2-benzothiazolyl)propionicacid dihydrochloride

(solid)

¹ -NMR (DMSO-d₆) δ:2.00 (2H, br), 2.26 (1.5H), 2.30 (1.5H), 3.00-4.25(7H), 5.17 (1H, br), 6.99 (2H, d), 7.31 (2H, d), 7.88 (1H, d), 8.25 (1H,d), 8.44 (1H, d), 8.50 (1H), 9.33 (2H, br), 9.55 (2H, br)

INVENTIVE EXAMPLE 902-[4-[((3R)-1-acetimidolyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-amidino-1-ethyl-2-indolyl)propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.31 (3H, t, J=7.0Hz), 2.15-2.40 (2H, m), 2.28(1.5H), 2.31 (1.5H), 3.15 (1H, dd), 3.40-4.05 (5H), 4.10 (1H, t), 4.28(2H, m), 5.16 (0.5H, br), 5.22 (0.5H, br), 6.40 (1H, s), 6.97 (2H, dd),7.40 (2H), 7.48 (1H, d, J=8.3Hz), 7.62 (1H, d, J=8.3Hz), 8.13 (1H, s),8.50 (0.5H, s), 8.59 (0.5H, s), 8.98 (2H, s), 9.32 (2H, s), 9.25-9.35(1H)

INVENTIVE EXAMPLE 912-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:2.00-2.40 (5H, m), 2.90-4.10 (7H, m), 5.15 (1H, br),6.93 (2H, d, J=8.0Hz), 7.33 (2H, d, J=8.0Hz), 7.50-8.40 (6H, m),8.50-8.70 (1H), 9.30 (3H, br), 9.55 (2H, br)

INVENTIVE EXAMPLE 922-[4-[((2R)-1-acetimidoyl-2-pyrrolidinyl)methoxy]phenyl]-3-(5-amidinobenzo[b]thien-2-yl)propionicacid dihydrochloride

1.0 g of ethyl3-(5-amidinobenzo[b]thien-2-yl)-2-ethoxycarbonyl-2-[4-[((2R)-2-pyrrolidinyl)methoxy]phenyl]propionatedihydrochloride was dissolved in 20 ml of ethanol, followed by theaddition of 0.42 g of ethyl acetimidate hydrochloride. 0.51 g oftriethylamine was added to the thus prepared solution during stirringunder ice cooling, and the resulting mixture was warmed up to roomtemperature and stirred for 18 hours. Thereafter, the solvent wasdistilled off to obtain ethyl2-[4-[((2R)-1-acetimidoyl-2-pyrrolidinyl)methoxy]phenyl]-2-ethoxycarbonyl-3-(5-amidinobenzo[b]thien-2-yl)propionatedihydrochloride. The thus obtained ester compound was dissolved in 50 mlof 5N hydrochloric acid and refluxed under heating for 60 minutes. Afterdistilling off the solvent, the resulting residue was subjected tocolumn chromatography using a column packed with a highly porous polymertype synthetic adsorbent (styrene-divinylbenzene polymer: Diaion HP-20)and using a water/acetonitrile mixture as an elution solvent. Fractionsof interest were pooled and concentrated, and the resulting residue wassubjected to reversed phase high performance liquid chromatography usinga column packed with octadecyl-bonded silica gel and using awater/acetonitrile mixture as an elution solvent. Thereafter, the thuseluted fractions of interest were pooled, mixed with dilute hydrochloricacid, and then concentrated to dryness. In this way, 360 mg of the titlecompound was obtained in the form of a light yellow solid.

¹ H-NMR (DMSO-d₆) δ:2.00 (4H, br), 2.24-2.43 (3H), 3.00-4.00 (5H), 4.00(2H), 4.50 (1H, br), 6.90 (2H, d), 7.30 (3H), 7.70 (1H, d), 8.10 (1H,d), 8.23 (1H, s), 9.20-9.60 (5H, m)

INVENTIVE EXAMPLE 933-(5-amidinobenzoyl[b]thien-2-yl)-2-[4-[((3S)-1-benzimidoyl-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

1.0 g of ethyl3-(5-amidinobenzo[b]thien-2-yl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride was dissolved in 15 ml of ethanol. To this solution wasadded 773 mg of ethyl benzimidate hydrochloride which has been preparedby allowing benzonitrile to react with ethanol in the presence ofhydrogen chloride. 631 mg of triethylamine was added to the thusprepared solution during stirring under ice cooling, and the mixture waswarmed up to room temperature, and stirred for 18 hours. Thereafter, thesolvent was distilled off to obtain ethyl3-(5-amidinobenzo[b]thien-2-yl)-2-[4-[((3S)-1-benzimidoyl-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride. The thus obtained ester compound was dissolved in 60 mlof 3N hydrochloric acid and refluxed under heating for 30 minutes. Afterdistilling off the solvent, the resulting residue was subjected tocolumn chromatography using a column packed with a highly porous polymertype synthetic adsorbent (styrene-divinylbenzene polymer: Diaion HP-20)and using a water/acetonitrile mixture as an elution solvent. Fractionsof interest were pooled and concentrated, and the resulting residue wassubjected to reversed phase high performance liquid chromatography usinga column packed with octadecyl-bonded silica gel and using awater/acetonitrile mixture as an elution solvent. Thereafter, the thuseluted fractions of interest were pooled, mixed with dilute hydrochloricacid, and then concentrated to dryness. In this way, 350 mg of the titlecompound was obtained in the form of a light yellow solid.

¹ H-NMR (DMSO-d₆) δ:2.00-2.80 (2H, m), 3.00-3.30 (7H, m), 4.04 (0.5H,br), 4.30 (0.5H, br), 6.80-7.90 (11H, m), 8.12 (1H, d, J=8.3Hz), 8.30(1H, s), 9.20-9.70 (6H, m)

The following compounds of Inventive Examples 94 to 100 were prepared inaccordance with the procedure of Inventive Example 93.

INVENTIVE EXAMPLE 943-(5-amidinobenzo[b]thien-2-yl)-2-[4-[((3S)-1-n-hexan-imidoyl-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:0.80-0.95 (3H, m), 1.20-1.40 (4H, m), 1.45-1.70 (2H,m), 2.15-2.40 (2H, m), 2.45-2.60 (2H, m), 3.25-3.90 (6H, m), 3.96 (1H,t, J=7.5Hz), 6.85-7.00 (2H, m), 7.25-7.40 (3H, m), 7.69 (1H, dd, J=8.3and 1.5Hz), 8.11 (1H, d, J=8.3Hz), 8.25 (1H, s), 8.58 (0.5H, s), 8.66(0.5H, s), 9.20-9.30 (3H, br), 9.47 (2H, br)

INVENTIVE EXAMPLE. 953-(5-amidinobenzo[b]thien-2-yl)-2-[4-[((3S)-1-cyclopropanecarboximidoyl-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:0.90-1.30 (3H, m), 1.80-4.10 (10H, m), 5.10-5.30(1H, m), 6.96 (2H, d, J=8.4Hz), 7.32 (1H, s), 7.36 (2H, d, J=8.4Hz),7.71 (1H, d, J=7.4Hz), 8.14 (1H, d, J=7.4Hz), 8.29 (1H, 8.40-8.70 (2H,m), 9.36 (2H, br), 9.52 (2H, br)

INVENTIVE EXAMPLE 962-[-4-[((2S)-1-acetimidoyl-2-pyrrolidinyl]methoxy]phenyl]-3-(5-amidinobenzo[b]thien-2-yl)propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:2.05 (4H, br), 2.25-2.43 (3H), 3.00-4.50 (8H), 6.95(2H), 7.30 (3H), 7.70 (1H, d), 8.10 (1H, d), 8.25 (1H, s), 8.60 (1H, s),9.20-9.60 (5H, m)

INVENTIVE EXAMPLE 972-[4-[((1-acetimidoyl-4-piperidinyl)oxy]phenyl]-3-(5-amidinobenzo[b]thien-2-yl)propionic acid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.65-2.10 (4H, m), 2.32 (3H, s), 3.20-4.00 (7H, m),4.60-4.70 (1H, m), 6.96 (2H, d, J=8.3Hz), 7.30 (3H, m), 7.69 (1H, d,J=8.3Hz), 8.10 (1H, d, J=8.3Hz), 8.26 (1H, s), 8.95 (1H, s), 9.32 (2H,br), 9.52 (2H, br)

INVENTIVE EXAMPLE 982-[4-[(1-acetimidoyl-4-piperidinyl)oxy]phenyl]-3-(6-amidino-1-ethyl-2-indolyl)propionicacid dihydrochloride

¹ H-NMR (DMSO-d₆) δ:1.30 (3H, t, J=7.0Hz), 1.73-2.10 (4H, m), 2.31 (3H,s), 3.10 (1H, m), 3.30-3.80 (5H), 4.05 (1H, t), 4.30 (2H, m), 4.70 (1H,br), 6.38 (1H, s), 6.97 (2H, d, J=8.5Hz), 7.37 (2H, d, J=8.5Hz), 7.48(1H, d, J=8.3Hz), 7.61 (1H, d, J=8.3Hz), 8.14 (1H, s), 8.86 (1H, br),9.15-9.50 (5H, m)

INVENTIVE EXAMPLE 993-(7-amidino-2-naphthyl)-2-[4-[((3S)-1-butanimidoyl-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ: 0.60-4.00 (16H), 5.00 (1H, br), 6.79 (2H, d,J=8.0Hz), 7.21 (2H, d, J=8.0Hz), 7.30-8.10 (5H, m), 8.34 (1H, s), 8.30(1H, s), 8.40-8.70 (1H), 9.00-10.00 (5H)

INVENTIVE EXAMPLE 1003-(7-amidino-2-naphthyl)-2-[4-[((3S)-1-benzimidoyl-3-pyrrolidinyl)oxy]phenyl]propionateacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:2.00-4.10 (9H), 4.90 (0.5H, br), 5.20 (0.5H, br),6.70-8.10 (14H, m), 8.32 (1H, s), 9.10-9.50 (4H)

INVENTIVE EXAMPLE 1013-(5-amidinobenzo[b]thien-2-yl)-2-[4-[[(3S)-1-(N-methyl-acetimidoyl)-3-pyrrolidinyl]oxy]phenyl]propionicacid dihydrochloride

2.0 g of3-(5-amidinobenzo[b]thien-2-yl)-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionicacid dihydrochloride was dissolved in a solvent mixture of 10 ml ofwater and 10 ml of acetonitrile. With stirring, to the thus preparedsolution was gradually added 20 g of ethyl (N-methyl)acetimidatehydrochloride which has been prepared in accordance with the proceduredisclosed in The Journal of Organic Chemistry (vol.33, pp.1679-1681,1968), while maintaining the pH level of the solution at 8.5 with 2Nsodium hydroxide aqueous solution. After distilling off the solvent, theresulting residue was washed with dichloromethane and then subjected tocolumn chromatography using a column packed with a highly porous polymertype synthetic adsorbent (styrene-divinylbenzene polymer: Diaion HP-20)and using a water/acetonitrile mixture as an elution solvent, therebyeffecting desalting. Fractions of interest thus pooled were subjected toreversed phase high performance liquid chromatography using a columnpacked with octadecyl-bonded silica gel. Thereafter, the thus elutedfractions of interest were pooled and then concentrated to dryness bypassing through a C1 type of strongly basic ion exchange resin (DiaionSA-10, Nippon Rensui Co., Ltd.). In this way, 370 mg of the titlecompound was obtained in a solid form.

¹ H-NMR (DMSO-d₆) δ:2.00-2.44 (2H, m), 2.30 (1.5H), 2.33 (1.5H), 2.98(3H), 3.06-4.20 (7H, m), 5.00-5.40 (1H, br), 6.92 (2H, d, J=8.3Hz),7.l28 (1H, s), 7.33 (2H, d, J=8.3Hz), 7.72 (1H, d, J=9.01Hz), 8.06 (1H,d, J=9.0Hz), 8.28 (1H, s), 8.8-9.20 (1H, br), 9.23 (2H, br), 9.50 (2H,br)

INVENTIVE EXAMPLE 1023-(5-amidino-2-benzofuranyl)-2-[4-[((2R)-2-amino-1-butyl)oxy]phenyl]propionicacid dihydrochloride

In 300 ml of tetrahydrofuran were dissolved 1.1 g of ethyl3-(5-cyano-2-benzofuranyl)-2-(4-hydroxyphenyl)propionate, 1.24 g of(2R)-2-tert-butoxycarbonyl-amino-1-butanol and 1.72 g oftriphenylphosphine. The thus prepared solution was mixed with 1.14 g ofdiethyl azodicarboxylate and stirred at room temperature for 18 hours.The resulting solution was further mixed with 0.83 g of(2R)-2-tert-butoxycarbonylamino-1-butanol, 1.2 g of triphenylphosphineand 0.76 g of diethyl azodicarboxylate, and the mixture was stirred atroom temperature for 18 hours. After concentrating the thus preparedreaction solution to dryness, the resulting residue was purified bysubjecting it to silica gel column chromatography using a toluene/ethylacetate mixture as a developing solvent, thereby obtaining 660 mg ofethyl2-[4-[((2R)-2-tert-butoxycarbonylamino-1-butyl)oxy]phenyl]-3-(5-cyano-2-benzofuranyl)propionatein a colorless and oily form.

Thereafter, 660 mg of ethyl2-[4-[((2R)-2-tert-butoxycarbonylamino-1-butyl)oxy]phenyl]-3-(5-cyano-2-benzofuranyl)propionateobtained above was treated and purified in the same manner as describedin Inventive Example 50 to obtain 78 mg of the title compound in a solidform.

¹ H-NMR (DMSO-d₆) δ:1.04 (3H, t), 1.70 (2H), 3.0-4.2 (6H), 6.71 (1H, s),6.99 (2H, d),l 7.27 (2H, d), 7.72 (2H, s), 8.07 (1H, s), 8.3 (3H, br),9.34-9.40 (4H)

INVENTIVE EXAMPLE 1033-(5-amidino-2-benzofuranyl)-2-[4-[((2S)2-amino-1-butyl)oxy]phenyl]propionateacid dihydrochloride

This compound was obtained in a solid form with a yield of 620 mg byrepeating the process of Inventive Example 102, except that(2S)-2-tert-butoxycarbonylamino-1-butanol was used instead of(2R)-2-tert-butoxycarbonylamino-1-butanol.

¹ H-NMR (DMSO-d₆) δ:1.04 (3H, t), 1.70 (2H), 3.0-4.2 (6H), 6.68 (1H, s),6.96 (2H, d), 7.24 (2H, d), 7.70 (2H, s), 8.05 (1H, s), 8.4 (3H, br),9.40 (4H, br)

INVENTIVE EXAMPLE 1043-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-4-(5-amidinobenzo[b]thien-2-yl)butyricacid dihydrochloride

1 ml of thionyl chloride was added dropwise to 50 ml of ethanol. Withstirring at room temperature, to this solution was added 1.0 g of4-(5-amidinobenzo[b]thien-2-yl)-3-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]butyricdihydrochloride, followed by refluxing under heating for 1 hour. Aftercooling and subsequent removal of the solvent by distillation, the thusprepared reaction solution was thoroughly dried under a reduced pressureto obtain ethyl4-(5-amidinobenzo[b]thien-2-yl)-3-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]butyratedihydrochloride. The thus obtained ester compound was dissolved in 20 mlof tetrahydrofuran. During ice cooling and stirring, the thus preparedsolution was mixed with triethylamine and then with 360 mg of ethylacetimidate hydrochloride, and the resulting mixture was stirred for 1hour. After distilling off the solvent, the resulting residue wassubjected to column chromatography using a column packed with a highlyporous polymer type synthetic adsorbent (styrene-divinylbenzene polymer:Diaion HP-20) and using a water/acetonitrile mixture as an elutionsolvent. Fractions of interest were pooled and concentrated to dryness,and the resulting residue was dissolved in 50 ml of 2N hydrochloric acidand refluxed under heating for 1 hour. After distilling off the solvent,the resulting residue was subjected to column chromatography using acolumn packed with a highly porous polymer type synthetic adsorbent(styrene-divinylbenzene polymer: Diaion HP-20) and using awater/acetonitrile mixture as an elution solvent. The fractions ofinterest were pooled and subjected to reversed phase high performanceliquid chromatography using a column packed with octadecyl-bonded silicagel and using a water/acetonitrile mixture as an elution solvent.Thereafter, the thus eluted fractions of interest were pooled, mixedwith dilute hydrochloric acid, and then concentrated to dryness. In thisway, 850 mg of the title compound was obtained in a solid form.

¹ H-NMR (DMSO-d₆) δ:2.0-2.45l(2H, m), 2.32 (3H, d), 2.5-2.9 (2H, m),3.1-4.0 (7H, m), 5.1-5.35 (1H, m), 6.92 (2H, d), 7.30 (2H, d), 7.8 (1H,d), 8.20 (1H, d), 8.37 (1H, s), 8.6-8.9 (1H, m), 9.30-9.80 (5H)

FAB MS (m/z): 465

INVENTIVE EXAMPLE 1052-[4-[((3S)-1-(acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(6-amidino-1-ethyl-2-indolyl)propionicacid dihydrochloride

This compound was prepared in accordance with the procedure of InventiveExample 104.

(solid)

¹ H-NMR (DMSO-d₆) δ:1.35 (3H, t), 2.37 (3H), 4.10-4.40 (1H, m), 6.42(1H, s), 7.00 (2H, a), 7.45 (2H, d), 7.60 (2H, m), 8.30 (1H, s), 8.70(1H, br), 9.25-9.80 (5H)

INVENTIVE EXAMPLE 1063-[4-[((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-2-(5-amidinobenzo[b]thien-2-yl)propionicacid dihydrochloride

2.0 g of ethyl3-[4-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]phenyl]-2-(5-cyanobenzo[b]thien-2-yl)propionatewas dissolved in 100 ml of ethanol. With ice cooling and stirring,hydrogen chloride was bubbled into the thus prepared solution to asaturation level. The resulting solution was allowed to stand still atroom temperature for 18 hours. After distilling off the solvent, thethus obtained residue was dissolved in 100 ml of ethanol solutioncontaining 13% ammonia, and the solution was allowed to stand still for18 hours. After distilling off the solvent, the resulting residue wassubjected to column chromatography using a column packed with a highlyporous polymer type synthetic adsorbent (styrene-divinylbenzene polymer:Diaion HP-20) and using a water/acetonitrile mixture as an elutionsolvent, thereby obtaining 1.1 g of ethyl2-(5-amidinobenzo[b]thien-2-yl)-3-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propionatedihydrochloride. 1.1 g of the i thus obtained compound was dissolved in15 ml of ethanol, and the solution was mixed with 566 mg of ethylacetimidate hydrochloride and 694 mg of triethylamine in that order,followed by 18 hours of stirring at room temperature. After distillingoff the solvent, the thus obtained residue was dissolved in 50 ml of 2Nhydrochloric acid, and then refluxed under heating for 30 minutes. Aftercooling and subsequently distilling off the solvent, the resultingresidue was subjected to column chromatography using a column packedwith a highly porous polymer type synthetic adsorbent(styrene-divinylbenzene polymer: Diaion HP-20) and using awater/acetonitrile mixture as an elution solvent. Fractions of interestthus pooled were subjected to reversed phase high performance liquidchromatography using a column packed with octadecyl-bonded silica geland using a water/acetonitrile mixture as an elution solvent.Thereafter, the thus eluted fractions of interest were pooled, mixedwith dilute hydrochloric acid, and then concentrated to dryness. In thisway, 490 mg of the title compound was obtained in a solid form.

¹ H-NMR (DMSO-d₆) δ:2.1-2.4 (2H, m), 2.22 (1.5H), 2.29 (1.5H), 3.08 (1H,dd, J=13.7 and 7.8Hz), 3.30-4.00 (5H, m), 4.36 (1H), 5.00-5.20 (1H),6.80-6.90 (2H, m), 7.15-7.25 (2H, m), 7.44 (1H, s), 7.72 (1H, d,J=8.3Hz), 8.15 (1H, d, J=8.3Hz), 8.32 (1H, s), 8.58 (0.5H), 8.66 (0.5H),9.32 (2H, br), 9.38 (0.5H), 9.45 (0.5H), 9.50 (2H, br)

The following compounds of Inventive Examples 107 to 110 were preparedin accordance with the procedure of Inventive Example 106.

INVENTIVE EXAMPLE 1072-[4-[((3R)-1-acetimidoyl-3-pyrrolidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:2.00-2.40 (5H, m), 2.90-4.10 (7H, m), 5.20 (1H, br),6.93 (2H, d, J=8.0Hz), 7.33 (2H, d, J=8.0Hz), 7.56 (1H, d), 7.70l-8.20(4H, m), 8.45 (1H, s), 8.50-8.80 (1H), 9.45 (3H, br), 9.63 (2H, br)

INVENTIVE EXAMPLE 1082-[4-[(1-acetimidoyl-4-piperidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.50-2.10 (4H, m), 2.31 (3H, s), 3.00-4.20 (7H, m),4.60-4.80 (1H, m), 6.95 (2H, d, J=9.0Hz), 7.31 (2H, d, J=9.0Hz),7.50-8.50 (6H, m), 8.93 (1H), 9.45 (3H, br), 9.62 (2H, br)

INVENTIVE EXAMPLE 1093-(7-amidino-2-naphthyl)-2-[4-[(1-butanimidoyl-4-piperidinyl)oxy]phenyl]propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:0.96 (3H, t, J=7.0Hz), 1.57 (2H, m), 1.71 (2H, br),2.03 (2H, br), 2.52 (2H, t), 3.18 (1H, m), 3.40-3.90 (5H), 4.00 (1H, t),4.68 (1H, br), 6.96 (2H, d, J=8.0Hz), 7.30 (2H, d, J=8.0Hz), 7.60-8.40(6H, m), 8.86 (1H), 9.32 (3H, br), 9.58 (2H, br)

INVENTIVE EXAMPLE 1102-[4-[((2R,4S)-1-acetimidoyl-2-methyl-4-pyrrolidinyl)oxy]phenyl]-3-(5-amidinobenzo[b]thien-2-yl)propionicacid dihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.35 (3H, m), 2.27 (1.5H, s), 2.37 (1.5H, s),5.10-5.50 (1H, br), 7.00 (2H, d), 7.10-8.70 (6H, m), 9.10-9.60 (6H, br)

FAB MS (m/z): 465 (M⁺ +1)

INVENTIVE EXAMPLE 1113-(5-amidinobenzo[b]thien-2-yl)-2-[4-[((2S)-5-oxo-2-pyrrolidinyl)methoxy]phenyl]propionicacid hydrochloride

a) 3.2 g of ethyl2-[4-[((2S)-1-tert-butoxy-carbonyl-5-oxo-2-pyrrolidinyl)methoxy]phenyl]-3-(5-cyanobenzo[b]thien-2-yl)propionatewas dissolved in a solvent mixture of 50 ml of ethanol and 50 ml ofdichloromethane. During ice-cooling and stirring, hydrogen chloride wasbubbled into this solution to a saturation level. The thus preparedreaction mixture was allowed to stand still at 5° C. for 48 hours. Afterdistilling off the solvent, the thus obtained residue was dissolved in100 ml of ethanol solution containing 13% (w/v) of ammonia, and thesolution was maintained at room temperature for 24 hours. Thereafter,the solvent was removed by distillation to obtain ethyl3-(5-amidinobenzo[b]thien-2-yl)-2-[4-[((2S)-5-oxo-2-pyrrolidinyl)methoxy]phenyl]propionatehydrochloride. The thus obtained ester compound was dissolved in asolvent mixture of 30 ml of tetrahydrofuran and 30 ml of water. To theresulting mixture was added 1.6 g of2-(tert-butoxycarbonylimino)-2-phenylacetonitrile and 2 ml of1,8-diazabicyclo[5.4.0]-7-undecene. After stirring at room temperaturefor 24 hours, the resulting reaction solution was extracted with ethylacetate and dried. After distilling off the solvent, the thus obtainedresidue was purified by subjecting it to silica gel columnchromatography using a chloroform/methanol mixture as an eluant, therebyobtaining 2.4 g of ethyl3-[5-(N-tert-butoxycarbonyl)aminoiminomethylbenzo[b]thien-2-yl]-2-[4-[((2S)-5-oxo-2-pyrrolidinyl)methoxy]phenyl]propionatein a solid form.

¹ H-NMR (CDCl₃) δ:1.18 (3H, t), 1.58 (9H, s), 1.80-2.50 (4H, m), 3.28(1H, dd), 3.70 (1H, dd), 3.80-4.50 (6H, m)

b) 2.4 g of ethyl3-[5-(N-tert-butoxycarbonyl)aminoiminomethylbenzo[b]thien-2-yl]-2-[4-[((2S)-5-oxo-2-pyrrolidinyl)methoxy]phenyl]propionateobtained in the above step a) was dissolved in 30 ml of tetrahydrofuran.The thus prepared solution was mixed with an aqueous solution of 200 mgof sodium hydroxide dissolved in 5 ml of water, and the mixture wasstirred for 72 hours. After distilling off the solvent, the thusobtained residue was dissolved in 10 ml of concentrated hydrochloricacid, and the solution was stirred at room temperature for 1 hour. Afterdistilling off the solvent, the resulting residue was subjected tocolumn chromatography using a column packed with a highly porous polymertype synthetic adsorbent (styrene-divinylbenzene polymer: Diaion HP-20)and using a water/acetonitrile mixture as an elution solvent.Thereafter, thus eluted fractions of interest were pooled, mixed withhydrochloric acid, and then concentrated to dryness. In this way, 1.1 gof the title compound was obtained.

¹ H-NMR (DMSO-d₆) δ:1.70-2.30 (4H, m), 6.90 (2H, d), 7.29 (2H, d), 7.30(1H, s)

FAB MS (m/z): 438 (M⁺ +1)

INVENTIVE EXAMPLE 1122-[2-[4-[(4-imidazolyl)methoxy]phenyl]ethyl]-5-benzofurancarboxamidinedihydrochloride

3.53 g portion of2-[2-[4-[(1-trityl-4-imidazolyl)methoxy]phenyl]ethyl]-5-benzofurancarbonitrilewas dissolved in a solvent mixture of 150 ml of ethanol and 100 ml ofdichloromethane. Hydrogen chloride acid gas was bubbled into the thusprepared solution while stirring under ice cooling, and the thussaturated solution was maintained at room temperature for 24 hours.After distilling off the solvent, the thus obtained residue wasdissolved in an ethanol solution containing 15% (w/v) of ammonia, andthe solution was stirred at room temperature for 80 hours. Afterdistilling off the solvent, the resulting residue was dissolved in amixture of 100 ml of formic acid and 2 ml of concentrated hydrochloricacid, and the thus prepared solution was stirred for 6 hours. Afterremoving formic acid by distillation, the thus obtained residue wasdissolved in hot water, and insoluble materials were removed byfiltration. After concentrating the thus obtained filtrate to dryness,the resulting residue was subjected to column chromatography using acolumn packed with a highly porous polymer type synthetic adsorbent(styrene-divinylbenzene polymer: Diaion HP-20) and using awater/acetonitrile mixture as an elution solvent. Fractions of interestwere pooled and concentrated, and the resulting residue was subjected toreversed phase high performance liquid chromatography using a columnpacked with octadecyl-bonded silica gel and using a water/acetonitrilemixture as an elution solvent. Thereafter, the thus eluted fractions ofinterest were pooled, mixed with dilute hydrochloric acid, and thenconcentrated to dryness. In this way, 730 mg of the title compound wasobtained in the form of a light yellow solid.

¹ H-NMR (DMSO-d₆) δ: 2.95 (4H, m), 5.12 (2H, s), 6.72 (1H, s), 6.94 (2H,d, J=8.7Hz), 7.20 (2H, d, J=8.7Hz), 7.72 (2H, s), 7.75 (1H, s), 8.08(1H, s), 9.13 (3H, br), 9.38 (2H, br)

INVENTIVE EXAMPLE 1132-[2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]ethyl]-6-indolecarboxamidinedihydrochloride

650 mg of2-[2-[4-[((3S)-1-tert-butoxycarbonyl-3pyrrolidinyl)oxy]phenyl]ethyl]-6-indolecarbonitrilewas dissolved in a solvent mixture of 100 ml of ethanol and 30 ml ofdichloromethane. During ice cooling and stirring, hydrogen chloride wasbubbled into the thus prepared solution to a saturation level. Theresulting solution was allowed to stand still at room temperature for 24hours. After distilling off the solvent, the resulting residue wasdissolved in an ethanol solution containing 11% (w/v) of ammonia, andthe solution was stirred at room temperature for 24 hours. Afterdistilling off the solvent, the resulting residue was subjected toreversed phase high performance liquid chromatography using a columnpacked with octadecyl-bonded silica gel and using a water/acetonitrilemixture as an elution solvent. Thereafter, the thus eluted fractions ofinterest were pooled, mixed with dilute hydrochloric acid, and thenconcentrated to dryness. In this way, 90 mg of the title compound wasobtained in the form of crystals.

mp: 229° to 233° C.

¹ H-NMR (DMSO-d₆) δ:1.95-2.35 (2H, m), 5.00-5.30 (1H, br), 6.36 (1H, s),6.80-7.80 (7H, m), 8.00 (1H, s), 9.30-9.60 (6H, br)

The following compounds of Inventive Examples 114 to 121 were preparedin accordance with the procedure of Inventive Example 113.

INVENTIVE EXAMPLE 1142-[3-hydroxy-2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propyl]-6-indolecarboxamidinedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:1.97-2.30 (2H, m), 2.90-4.60 (9H, m), 5.00-5.20 (1H,br), 6.22 (1H, s), 6.90 (2H, d), 7.18-7.70 (2H, m), 7.96 (1H, s),9.10-9.90 (6H, br), 11.05 (1H, s)

INVENTIVE EXAMPLE 115 2-[2-[4-[[((2-pyrazinyl)amino]carbonyl]phenyl]ethyl]-5-benzofurancarboxamidinedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:3.20 (4H, 6.78 (1H, s), 7.08 (1H, br), 7.48 (2H, d,J=7.9Hz), 7.80 (2H, s), 8.03 (2H, d, J =7.9Hz), 8.12 (1H, s), 8.40-8.60(2H, m), 9.25 (2H, br), 9.39 (3H, br), 11.05 (1H, s)

INVENTIVE EXAMPLE 1162-[2-[4-[(1-imidazolyl)methyl]phenyl]ethyl]-5-benzofurancarboxamidinedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ: 3.10 (4H, s), 5.45 (2H, s), 6.72 (1H, s), 7.24 (2H,d, J=8.3Hz), 7.40 (2H, d, J=8.3Hz), 7.66 (1H, s), 7.72 (2H, s), 7.80(1H, s), 8.12 (1H, s), 9.30 (2H, br), 9.45 (3H, br)

INVENTIVE EXAMPLE 1172-[2-[4-[(4-methyl-1-piperazinyl)carbonyl]phenyl]ethyl]-5-benzofurancarboxamidinedihydrochloride

¹ H-NMR (DMSO-d₆) δ: 2.80 (3H, s), 4.09 (4H, s), 3.10 (4H, br), 4.00(4H, br), 6.74 (1H, s), 7.36 (4H, s), 7.74 (2H, s), 8.12 (1H, s), 9.28(2H, br), 9.48 (2H, br)

INVENTIVE EXAMPLE 1183-[3-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]propyl]-5-benzofurancarboxamidinedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ: 2.10 (4H, m), 2.70 (4H, m), 3.30 (4H), 5.07 (1H,br), 6.90 (2H, d), 7.15 (2H, d), 7.79 (2H, s), 8.00 (1H, s), 8.23 (1H,s), 9.20-9.80 (6H, br)

INVENTIVE EXAMPLE 1192-[[4-[(4-piperidinyl)oxy]phenyl]methyl]-5-benzofurancarboxamidinedihydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ: 1.70-2.20 (2H, m), 2.70-3.30 (4H, m), 4.14 (2H, s),4.60-4.80 (1H, m), 6.79 (1H, s), 6.97 (2H, d, J=9.0Hz), 7.26 (2H, d,J=9.0Hz), 7.74 (2H, s), 8.13 (1H, s), 9.30 (2H, br), 9.44 (2H, br),9.00-9.60 (2H, br)

INVENTIVE EXAMPLE 1202-[2-[4-[((3S)-1-acetyl-3-pyrrolidinyl)oxy]-3-hydroxyphenyl]ethyl]-5-benzofurancarboxamidinehydrochloride

mp: 175°-176° C.

¹ H-NMR (DMSO-d₆) δ:1.80-2.20 (5H, m), 2.70-3.80 (8H, m), 4.88 (1H, br),6.60 (1H, d, J=8.3Hz), 6.77 (1H, s), 6.82 (1H, d, J=8.3Hz), 7.72 (1H, d,J=8.1Hz), 7.80 (1H, d, J=8.1Hz), 8.08 (1H, s), 8,90-8.98 (1H), 9.23 (2H,br), 9.40 (2H, br)

INVENTIVE EXAMPLE 1212-[2-[4-[(4-(N-acetyl)aminomethylcyclohexyl)methoxy]phenyl]ethyl]-5-benzofurancarboxamidinehydrochloride

(solid)

¹ H-NMR (DMSO-d₆) δ:0.70-2.00 (10H, m), 1.83 (3H, s), 2.75-3.20 (6H, m),3.70 (2H, d, J=5.7Hz), 6.64 (1H, s), 6.77 (2H, d, J=8.8Hz), 7.15 (2H, d,J=8.8Hz), 7.65-7.68 (3H), 8.04 (1H, s), 9.00 (2H, br), 9.31 (2H, br)

INVENTIVE EXAMPLE 1222-[2-[4-[((3S)-1-formimidoyl-3-pyrrolidinyl)oxy]phenyl]ethyl]-5-benzofurancarboxamidinedihydrochloride

a) The procedure of Inventive Example 100 was followed to synthesize2-[2-[4-[((3S)-1-tert-butoxy-carbonyl-3-pyrrolidinyl)oxy]phenyl]ethyl]-5-benzofurancarbonitrile.

¹ H-NMR (CDCl₃) δ: 1.66 (9H, s), 3.04 (4H, s), 3.30-3.70 (4H, br), 4.85(1H), 6.40 (1H, s), 6.80 (2H, d), 7.12 (2H, d), 7.52 (2H, s), 7.82 (1H,s)

b) 1.66 g of2-[2-[4-[((3S)-1-tert-butoxy-carbonyl-3-pyrrolidinyl)oxy]phenyl]ethyl]-5-benzofurancarbonitrileobtained in the above step a) was treated and purified in accordancewith the procedure of Inventive Example 113, thereby obtaining 800 mg of2-[2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]ethyl]-5-benzofurancarboxamidinedihydrochloride.

¹ H-NMR (DMSO-d₆) δ:1.90-2.30 (2H, m), 3.06 (4H, br), 3.00-3.80 (4H,br), 5.08 (1H, br), 6.73 (1H, s), 6.88 (2H, d), 7.19 (2H, d), 7.74 (2H,s), 8.11 (1H, s), 9.26 (2H, br), 9.47 (2H, br)

c) 1.0 g of2-[2-[4-[((3S)-3-pyrrolidinyl)oxy]phenyl]ethyl]-5-benzofurancarboxamidinedihydrochlorideobtained in the above step b) was dissolved in 15 ml of water. Duringice cooling and stirring, 1.83 g of benzyl methaneimidate hydrochloridewas added to the above solution while maintaining the reaction solutionat pH 8 with 1N sodium hydroxide aqueous solution. The thus preparedmixture was stirred for 20 minutes with ice cooling on an ice bath. Theresulting reaction solution was adjusted to pH 2.0 with dilutehydrochloric acid, washed with diethyl ether, and then concentrated todryness. Thereafter, the resulting residue was subjected to columnchromatography using a column packed with a highly porous polymer typesynthetic adsorbent (styrene-divinylbenzene polymer: Diaion HP-20) andusing a water/acetonitrile mixture as an elution solvent. In this way,0.76 g of the title compound was obtained in a solid form.

¹ H-NMR (DMSO-d₆) δ: 1.80-2.60 (2H, m), 3.08 (4H, br), 3.20-4.00 (4H,br), 5.14 (1H, br), 6.80 (1H, s), 6.92 (2H, d), 7.25 (2H, d), 7.76 (1H,d), 7.86 (1H, d), 8.21 (1H, s), 8.40 (1H, br), 9.08 (1H, br), 9.18 (2H,br), 9.57 (3H, br)

INVENTIVE EXAMPLE 1232-[2-[3-hydroxy-4-[((3S)-3-pyrrolidinyl)oxy]phenyl]ethyl]-5-benzofurancarboxamidinedihydrochloride

1.0 g of2-[2-[4-[((3S)-1-acetyl-3-pyrrolidinyl)oxy]-3-hydroxyphenyl]ethyl]-5-benzofurancarboxamidinehydrochloride was dissolved in 30 ml of 6N hydrochloric acid, and thesolution was refluxed under heating for 4 hours. After distilling offthe solvent, the resulting residue was subjected to columnchromatography using a column packed with a highly porous polymer typesynthetic adsorbent (styrene-divinylbenzene polymer: Diaion HP-20) andusing a water/acetonitrile mixture as an elution solvent. In this way,320 mg of the title compound was obtained in a solid form.

(solid)

¹ H-NMR (DMSO-d₆) δ:1.90-2.20 (2H, m), 2.70-3.50 (8H, m), 5.08 (1H, br),6.66 (1H, dd, J=9.0 and 1.8Hz), 6.80 (2H, s), 6.94 (1H, s), 7.76 (2H,s), 8.12 (1H, s), 9.26 (2H, br), 9.44 (2H, br), 9.74 (2H, br)

INVENTIVE EXAMPLE 1242-[2-[4-[(4-aminomethylcyclohexyl)methoxy]phenyl]ethyl]-5-benzofurancarboxamidinedihydrochloride

This compound was obtained in similar manner to the procedure ofInventive Example 123.

(solid)

¹ H-NMR (DMSO-d₆) δ:0.80-2.00 (10H, m), 2.90-3.20 (4H, m), 3.73 (4H, m),6.72 (1H, s), 6.79 (2H, d, J=8.5Hz), 7.14 (2H, d, J=8.5Hz), 7.73 (2H,s), 8.07 (3H, br), 9.18 (2H, br), 9.38 (2H, br)

INVENTIVE EXAMPLE 125[2-[2-(5-amidino-2-benzofuranyl)ethyl]-5-[((3S)-3-pyrrolidinyl)oxy]phenyl]oxyaceticacid dihydrochloride

1.6 g of methyl[5-[((3S)-1-tert-butoxycarbonyl-3-pyrrolidinyl)oxy]-2-[2-(5-cyano-2-benzofuranyl)ethyl]phenyl]oxyacetatewas dissolved in 100 ml of ethanol. With ice cooling and stirring,hydrogen chloride was bubbled into the thus prepared solution to asaturation level. The thus treated solution was warmed up to roomtemperature and then maintained at this temperature for 18 hours. Afterdistilling off the solvent, the thus obtained residue was dissolved in20 ml of ethanol, and the solution was added dropwise to an aqueoussolution of 5N sodium hydroxide under stirring. After 10 minutes ofstirring, the resulting reaction solution was saturated with sodiumchloride, extracted with chloroform three times (200 ml, 100 ml and 100ml in that order), and then dried on a potassium carbonate/magnesiumsulfate mixture. After distilling off the solvent, the thus obtainedresidue was dissolved in 50 ml of ethanol. The resulting solution wasmixed with 1.0 g of ammonium chloride and then with 200 ml of ethanolsolution containing 12% ammonia, and the mixture was stirred for 96hours. After distilling off the solvent, the thus obtained residue wassubjected to reversed phase high performance liquid chromatography usinga column packed with octadecyl-bonded silica gel and using awater/acetonitrile mixture as an elution solvent. Thereafter, thuseluted fractions of interest were pooled, mixed with dilute hydrochloricacid, and then concentrated to dryness. In this way, 0.7 g of the titlecompound was obtained in a powdery form.

¹ H-NMR (DMSO-d₆) δ: 1.96-2.32 (2H, br), 2.80-3.60 (8H, br), 4.75 (2H,s), 5.10 (1H, br), 6.48 (1H, d, J=8.8Hz), 6.51 (1H, s), 6.76 (1H, s),7.54 (1H, d, J=8.8Hz), 7.74 (2H, s), 8.10 (1H, s), 9.27 (2H, br), 9.42(2H, br), 9.00-10.20 (2H, br)

INVENTIVE EXAMPLE 126 ethyl[2-[2-(5-amidino-2-benzofuranyl)ethyl]-5-[((3S)-3-pyrrolidinyl)oxy]phenyl]oxyacetatedihydrochloride

0.65 g of[2-[2-(5-amidino-2-benzofuranyl)ethyl]-5-[((3S)-3-pyrrolidinyl)oxy]phenyl]oxyaceticacid dihydrochloride was dissolved in 50 ml of ethanol. 0.2 ml ofthionyl chloride was added to the above solution with ice cooling, andthe resulting mixture was stirred at room temperature for 18 hours.Thereafter, the resulting reaction solution was concentrated to dryness,dissolved once in water, and then concentrated again to dryness. In thisway, 0.65 g of the title compound was obtained in a solid form.

¹ H-NMR (DMSO-d₆) δ:1.20 (3H, t), 2.11 (2H, br), 3.30 (4H, br), 4.20(2H, q), 4.84 (2H, s), 5.08 (1H, br), 6.53 (2H, s), 6.75 (1H, s), 7.10(1H, d), 7.72 (2H, s), 8.07 (1H, s), 9.18 (2H), 9.38 (2H), 9.66 (2H, br)

TEST EXAMPLE 1 Measurement of Solubility in Water

A fixed amount of water was added to a varying amounts of each sample,and the mixture was agitated on a shaker at 25° C. for 10 minutes. Theresults are shown in Table 1.

                  TABLE 1                                                         ______________________________________                                        Compound             Solubility in Water                                      ______________________________________                                        DABE.sup.*1           5 mg/1 ml or less                                       Compound of Inventive Example 19                                                                   450 mg/1 ml or more                                      Compound of Inventive Example 21                                                                   600 mg/1 ml or more                                      Compound of Inventive Example 25                                                                   500 mg/1 ml or more                                      Compound of Inventive Example 45                                                                   200 mg/1 ml or more                                      Compound of Inventive Example 73                                                                   450 mg/1 ml or more                                      Compound of Inventive Example 96                                                                   400 mg/1 ml or more                                      Compound of Inventive Example 104                                                                  450 mg/1 ml or more                                      Compound of Inventive Example 108                                                                  350 mg/1 ml or more                                      ______________________________________                                         .sup.*1 : DABE, 1,2bis(5-amidino-2-benzofuranyl)ethane                   

Thus, the compounds of the present invention are clearly more soluble inwater than the prior art.

TEST EXAMPLE 2 Measurement of Anticoagulant Activity

A blood plasma sample was prepared from human blood using a centrifuge.A 100 μl portion of the blood plasma preparation was added to 100 μl ofphysiological saline solution containing with or without a compound tobe tested, and the mixture was allowed to stand still at 37° C. for 2minutes. 100 μl of 0.02M calcium chloride solution which has beenincubated at 37° C. in advance was added to the mixture. Thereafter,clotting time was measured using CLOTEC (Sanko Junyaku Co., Ltd.). Aclotting time measured without adding a compound to be tested was usedas a control, and a concentration of the test compound which doubles thecontrol clotting time was calculated (hereinafter referred to as "CT2")as an index of the anticoagulant activity. Typical examples of theresults are shown in Table 2.

TEST EXAMPLE 3 Measurement of Activated Blood Coagulation Factor X (FXa)Inhibition Activity

A 180 μl portion of physiological saline solution containing a compoundto be tested was mixed with 200 μl of Tris-HCl buffer (pH 8.4) and 100μl of 1 mM S-2222 (KabiVitrum AB) aqueous solution, and the mixture wasincubated at 37° C. 20 μl of Tris-HCl buffered saline (pH 7.45)containing 0.6 unit/ml of human FXa was added to the mixture. After 15minutes of incubation at the same temperature, the reaction was stoppedby adding 100 μl of 60% acetic acid, and absorbance of the reactionmixture was measured. A reaction mixture with no addition of a testcompound was used as a blank, and another reaction mixture in which 60%acetic acid was added prior to the addition of FXa was used as acontrol. A concentration of each compound to be tested which inhibits50% of the FXa activity was calculated (hereinafter, referred to as"IC₅₀ ") as an index of the FXa inhibition activity. Typical examples ofthe results are shown in Table 2.

TEST EXAMPLE 4 Measurement of Thrombin Inhibition Activity

A 100 μl portion of Tris-HCl buffered saline (pH 7.45) (TBS) containing6 mg/ml of fibrinogen (Type 1, Daiichi Pure Chemicals Co., Ltd.) wasmixed with 100 μl of physiological saline solution. At 37° C., 100 μl ofTris-HCl buffered saline (pH 7.45) (TBS) containing varying amounts ofthrombin (for topical application use, Sankyo Co., Ltd.) was added tothe mixture prepared above to measure clotting time using CLOTEC (SankoJunyaku Co., Ltd.) and to prepare a calibration curve. Inhibition ratio(%) of each compound to be tested was obtained by measuring clottingtime using 100 μl of physiological saline solution to which eachcompound has been added. A concentration of each compound which inhibits50% of the thrombin activity was calculated based on the inhibitionpercentage (hereinafter, referred to as "IC₅₀ ") and concentration ofthe compound was determined as an index of the Anti-thrombin activity.Typical examples of the results are also shown in Table 2.

                  TABLE 2                                                         ______________________________________                                                      Anti-       FXa      Thrombin                                                 Coagulation Inhibition                                                                             Inhibition                                               CT2         IC.sub.50                                                                              IC.sub.50                                  Compound      (μM)     (μM)  (μM)                                    ______________________________________                                        DABE          1.6         0.095    5                                          Compound of Inventive                                                                       0.32        0.032    9.0                                        Example 41                                                                    Compound of Inventive                                                                       0.18        0.013    >400                                       Example 43                                                                    Compound of Inventive                                                                       0.49        0.041    >2000                                      Example 45                                                                    Compound of Inventive                                                                       3           0.36     50                                         Example 48                                                                    Compound of Inventive                                                                       1.45        0.17     190                                        Example 68                                                                    Compound of Inventive                                                                       5           0.6      >600                                       Example 89                                                                    Compound of Inventive                                                                       1.1         0.1      370                                        Example 94                                                                    Compound of Inventive                                                                       0.8         0.16     26                                         Example 95                                                                    Compound of Inventive                                                                       0.54        0.044    29                                         Example 96                                                                    Compound of Inventive                                                                       0.23        0.045    170                                        Example 97                                                                    Compound of Inventive                                                                       0.3         0.011    2.5                                        Example 98                                                                    Compound of Inventive                                                                       0.64        0.086    230                                        Example 104                                                                   Compound of Inventive                                                                       0.35        0.054    6.8                                        Example 105                                                                   Compound of Inventive                                                                       2.3         0.56     100                                        Example 106                                                                   Compound of Inventive                                                                       0.3         0.018    250                                        Example 108                                                                   ______________________________________                                    

As is evident from the results shown in Table 2, each compound of thepresent invention shows a strong anticoagulant activity through itsspecific Anti-FXa activity, in comparison with DABE which is generallyknown as an anticoagulant agent.

TEST EXAMPLE 5 Measurement of Anticoagulant Activity in OralAdministration

An aqueous solution of each compound to be tested was administeredorally to each male STD:Wistar rat individual under an anesthetic, at adose of 10 ml/kg body weight. Blood samples were collected periodicallyand blood plasma preparations were obtained from the samples to measureactivated partial thromboplastin time (APTT). In the same manner, APTTwas measured by administering pure water and used as a control. Atest/control APTT ratio was calculated and used as an index of theanticoagulant activity. Typical examples of the results are shown inTable 3.

                  TABLE 3                                                         ______________________________________                                                         Test/Control APTT Ratio                                      Compound (dose)  0.5 hr  1 hr    2 hr  4 hr                                   ______________________________________                                        Compound of Inventive                                                                          1.63    1.52    1.48  1.28                                   Example 45 (100 mg/kg)                                                        Compound of Inventive                                                                          1.46    1.42    1.41  1.18                                   Example 48 (100 mg/kg)                                                        Compound of Inventive                                                                          1.40    1.28    1.21  1.09                                   Example 61 (100 mg/kg)                                                        Compound of Inventive                                                                          1.24    1.22    1.17  1.14                                   Example 89 (100 mg/kg)                                                        Compound of Inventive                                                                          1.68    1.64    1.57  1.42                                   Example 96 (100 mg/kg)                                                        Compound of Inventive                                                                          4.07    3.96    3.37  2.19                                   Example 98 (100 mg/kg)                                                        Compound of Inventive                                                                          2.69    3.60    2.41  1.66                                   Example 105 (100 mg/kg)                                                       Compound of Inventive                                                                          2.12    2.18    1.69  1.39                                   Example 108 (100 mg/kg)                                                       ______________________________________                                    

As shown in Table 3, a prolongation effect on plasma clotting time wasobserved clearly by the oral administration of each compound of thepresent invention.

TEST EXAMPLE 6 Toxicity Test by Single Oral Administration to Rat

The compound of Inventive Example 45 was dissolved in distilled water,and administered orally to each of two six-week old male Slc:SD rats, ata dose of 2,000 mg/kg body weight. No mortal case was found during 14days of observation.

TEST EXAMPLE 7 Toxicity Test by Repeated Oral Administration to Rat

Each of the compounds of the present invention was dissolved indistilled water and administered orally to each of five five-week oldmale Slc:SD rats, at a dose of 800 mg/kg body weight. The oraladministration was carried out once a day and repeated for 10 days toobserve mortality. The results are =shown in Table 4.

                  TABLE 4                                                         ______________________________________                                                         Individual Numbers                                                                          Mortal                                         Compound         Administered  Case                                           ______________________________________                                        Compound of Inventive                                                                          5             0                                              Example 19                                                                    Compound of Inventive                                                                          5             0                                              Example 25                                                                    Compound of Inventive                                                                          5             0                                              Example 45                                                                    Compound of Inventive                                                                          5             0                                              Example 69                                                                    Compound of Inventive                                                                          5             0                                              Example 88                                                                    ______________________________________                                    

TEST EXAMPLE 8 Antithrombotic Effect by Oral Administration to Rat in anArteriovenous Shunt Model

Amtithrombotic effect by oral administration of the compound of thepresent invention was measured by slightly modifying the proceduredisclosed in Thrombosis Research, vol. 54, pp. 399-410, 1989.

A predetermined amount of a compound to be tested was dissolved inpurified water and administered orally to a male STD:Wistar rat, and therat was anesthetized 15 minutes after the administration. An artery clipwas attached to proximally the carotid artery of the thus anesthetizedrat to stop blood circulation and to insert and fix an end of a shuntfilled with physiological saline, while the other end of the shunt wasinserted into the jugular vein and fixed. In this instance, the shuntwas prepared by indwelling a copper wire (0.17 mm in diameter and 20 cmin length) in a polyethylene tube (Hibiki No. 5; 5/3 mm in outerdiameter and 21 cm in length) and connecting each end of the tube with apolyethylene tube (Hibiki No. 3; 1 mm in outer diameter and 3 cm inlength) using a 3 mm silicone tube. After 30 minutes of theadministration, the artery clip was removed to flow blood into theshunt. After 7 minutes of the blood recirculation, the copper wire waspulled out together with formed thrombus and washed with 10 ml ofphysiological saline. Thereafter, the amount of thrombus formed on thecopper wire was determined as protein in accordance with the proceduredisclosed in Journal of Biological Chemistry, vol. 193, pp. 265-275,1951. As a control, water was administered instead of the compound to betested to calculate thrombosis inhibition rate after administration ofthe compound. The results are shown in Table 5.

                  TABLE 5                                                         ______________________________________                                                                 Thrombus  Thrombosis                                                Dose      formed    inhibition                                 Compound       (mg/kg)   (μg)   (%)                                        ______________________________________                                        Water          --        890 ± 102                                                                            --                                         Compound of Example 45                                                                       10        585 ± 85.sup.*1                                                                      34                                                        30        356 ± 51.sup.*1                                                                      60                                         ______________________________________                                         mean ± S.E. (n = 6)                                                        .sup.*1 : p < 0.05 (per control)                                         

As shown in Table 5, a significant thrombosis inhibition effect wasobserved with oral administration.

While the invention has been described in detail and with reference tospecific embodiments thereof, it will be apparent to one skilled in theart that various changes and modifications can be made therein withoutdeparting from the spirit and scope thereof.

What is claimed is:
 1. An aromatic amidine derivative represented by thefollowing general formula (1) or a pharmaceutically acceptable saltthereof: ##STR14## wherein R¹ is a hydrogen atom or a alkoxy group; R²is a hydrogen atom, a alkyl group, a alkoxy group, a carboxyl group, analkoxycarbonyl group, a carboxyalkyl group or an alkoxycarbonylalkylgroup;R³ is a hydrogen atom, a carboxyl group, an alkoxycarbonyl group,a carboxyalkyl group, an alkoxycarbonylalkyl group, a carboxyalkoxygroup or an alkoxycarbonylalkoxy group; R⁴ is a hydrogen atom, ahydroxyl group, a alkyl group or a alkoxy group; n is an integer of 0 to4; A is an alkylene group having a carbon number of 1 to 4, which mayhave 1 or 2 substituents selected from the group consisting ofhydroxyalkyl, carboxyl, alkoxycarbonyl, carboxyalkyl andalkoxycarbonylalkyl; X is a single bond, an oxygen atom, a sulfur atomor a carbonyl group; Y is a piperidinyl group optionally substitutedwith a member selected from the group consisting of an alkyl group, analkanoyl group, a carbamoyl group, a mono- or dialkylcarbamoyl group, aformimidoyl group, an alkanoimidoyl group, a benzimidoyl group, acarboxyl group, an alkoxycarbonyl group, a carboxyalkyl group, analkylcarbonylalkyl group, an aminoalkyl group, an alkanoylamino group,an alkanoylaminoalkyl group, an imino group, and alkoxycarbonyliminogroup, the group represented by ##STR15## is a member selected from thegroup consisting of indolyl, benzofuranyl, benzothienyl, benzimidazolyl,benzoxazolyl, benzothiazolyl, naphthyl, tetrahydronaphthyl and indanyl;and said alkyl, alkoxy or akanoyl group has a carbon number of 1 to 6.2. The aromatic amidine derivative or salt thereof according to claim 1,wherein said group represented by ##STR16## is a member selected fromthe group consisting of indolyl, benzofuranyl, benzothienyl,benzimidazolyl, benzothiazolyl, naphthyl and tetrahydronaphthyl.
 3. Anaromatic amidine derivative,2-[4-[(1-acetimidoyl-4-piperidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid, or a salt thereof.
 4. An aromatic amidine derivative,(+)-2-[4-[(1-acetimidoyl-4-piperidinyl)oxy]phenyl]-3-(7-amidino-2-naphthyl)propionicacid, or a salt thereof.
 5. An anticoagulation composition comprising ananticoagulative effective amount of the compound of claim 1 or a saltthereof as an active ingredient and a pharmaceutically acceptablecarrier.
 6. An anticoagulation composition comprising an anticoagulativeeffective amount of the compound of claim 2 or a salt thereof as anactive ingredient and a pharmaceutically acceptable carrier.
 7. Ananticoagulation composition comprising an anticoagulative effectiveamount of the compound of claim 4 or a salt thereof as an activeingredient and a pharmaceutically acceptable carrier.